Nabl 102 PDF

NABL 102
NABL
NATIONAL ACCREDITATION
BOARD FOR TESTING AND
CALIBRATION LABORATORIES
SPECIFIC CRITERIA
for BIOLOGICAL TESTING
LABORATORIES
ISSUE NO : 03
ISSUE DATE: 27.06.2012
AMENDMENT NO : 00
AMENDMENT DATE: --
AMENDMENT SHEET
Sl
Page
No.
Clause
Date of
Amendment
No.
Amendment made
Reasons
Signature
QO
Signature
Director
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National Accreditation Board for Testing and Calibration Laboratories

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Specific Criteria for Biological Testing Laboratories

Issue Date: 27.06.2012
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CONTENTS
S. No.
Title
Page No.
Amendment Sheet
Contents
1.
Introduction and Scope of Document
2.
Personnel
3.
Accommodation and Environment
4.
Test Methods and Method Validation
14
5.
Equipment - Maintenance, Calibration and Performance
21
6.
Sampling
35
7.
Sample Handling
37
8.
Disposal of Contaminated Waste
39
9.
Assuring Quality of Test Results
40
10.
Reporting the Results
44
Appendix A Classes of Tests in Biological Discipline
47
Appendix B Glossary of Terms
55
Appendix C Method Validation
58
Appendix D Reference Culture Maintenance, Subculture and Storage
60
Appendix E Bio-safety Levels
63
Appendix F Uncertainty of Measurement
65
References
70
Composition of the Technical Committee
72

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1.
Introduction and Scope of Document
1.1.
The general criteria for laboratory accreditation are laid down in the international standard,
General Requirements for the competence of testing and calibration laboratories (ISO/IEC
17025:2005). Laboratories seeking accreditation must meet all of these requirements.
1.2.
This document describes specific requirements that a biological testing laboratory has to meet, in
addition to the requirements of ISO/IEC 17025: 2005.
1.3.
This criteria document provides extra information and interpretation on classes of test, personnel,
accommodation and environment, equipments, reference materials/cultures and other aspects of
laboratory management practices which are considered to be minimum standards for biological
testing laboratories being accredited against NABL Laboratory Accreditation Program.
1.4.
As majority of accredited biological testing laboratories are primarily involved in bacteriological

testing this document does have a bias towards these types of laboratories. However other
emerging areas of biological testing like molecular biology, GMO testing, medical devices,
toxicology, veterinary science, biochemistry and cell culture etc has given more emphasis in this
revised document.
1.5.
Based upon assessors and laboratories feedback received with regard to not having a consensus
and definitive methodology for measurement uncertainty
in biological testing pertaining to
quantitative measurements, this document has tried to provide a better insight in the
understanding of this concept.
1.6.
In addition to this document there is one supplementary criteria document named as NABL-114
Guidelines for Food Testing Laboratories, being applicable to biological testing laboratories
engaged in the testing of food.
1.7.
For the purpose of covering the activities pertaining to grant of accreditation, a group wise list of
biological tests for which NABL offers accreditation is given in Annexure A, under the heading of
Classes of Tests in Biological Discipline. The way of identifying these activities for the purpose of
accreditation is perhaps a convenient means of expressing an accredited laboratory capability

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2.
Personnel (ISO/IEC 17025 clause 5.2)

The staff in the accredited laboratories must be competent and experienced in the relevant
technical area covered by their scope of accreditation. In addition, all the staff of the laboratory
shall have a general awareness about the requirements embodied in ISO/IEC 17025:2005.
Further the person designated as Quality Manager, shall have a formal certified training on
ISO/IEC 17025:2005.
2.1
The minimum qualification for the technical staff in a biological testing laboratory shall be
graduate in biology/ microbiology/fisheries/food science/food technology/ Pharmaceutical
Sciences/biotechnology/ molecular biology/biochemistry/toxicology/veterinary science. Alternative
qualifications in biological sciences may meet requirements where staff has relevant experience
relating to the laboratory's scope of accreditation.
Staff should have a minimum of 1 year of work experience in similar area covered by the scope of
accreditation as proven by demonstrated competence on records. Freshers can be put under
training with adequate supervision.
2.2
Authorized signatory should fulfill either of the following requirements :

(a) Graduates in the relevant field or equivalent with five years experience in similar area
out of which at least two years experience should be at supervisory level.
(b) Postgraduate/higher degree in the relevant field or equivalent with a minimum of two
years supervisory level experience in the relevant scope of accreditation.
Biological testing laboratories carrying out testing of pathogens shall have a qualified
microbiologist (graduate/post graduate in Microbiology with the above-mentioned experience) as
the authorized signatory.
The competence of authorized signatory will be assessed during the assessment before being
approved by NABL.
2.3
The laboratory management shall ensure that all personnel have received adequate training for
ensuring competency for the performance of assigned task. Personnel may only perform tests on
samples if they are either recognized as competent to do so, or working under adequate
supervision.

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2.4
Technical competence of the personnel shall be monitored objectively with provision for retraining where necessary. Where a method or technique is not in regular use, verification of
personnel performance is necessary before the testing is undertaken. The critical interval
between performances of non-routine tests should be established and documented by the
laboratory.
2.5
In addition to test methods, in some cases, it may be more appropriate to relate competence to a
particular technique or instrument, for example use of approved biochemical (i.e. FDA, AOAC),
serological kits or microbial identification kits.

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3.
Accommodation and Environment (ISO/IEC 17025 Clause 5.3)

The requirements of accommodation and environmental conditions for biological testing vary
widely depending on the nature of the testing involved by the laboratory.
3.1
Irrespective of the kind of tests being performed in the laboratory, there must be adequate space
and storage facilities for carrying out the tests, recording of test data and report preparation etc.
Risk of cross contamination or mix ups must be avoided at each test stage activities as it may
interfere or compromise the integrity of the data.
3.2
Storage facilities of the laboratories must be sufficient enough to allow the sample retention and if
required segregation of samples for designated periods and provide conditions that maintain
sample integrity.
3.3
Laboratory testing areas should have appropriate lighting, ventilation, adequate bench space,
and freedom from dust and fumes, control of temperature and humidity. The extent to which
these environmental factors apply will vary according to the type and precision of the testing.
3.4
The layout of laboratory should be arranged in such a way so that the risks of cross contamination
can be reduced. This can be achieved by carrying out the test procedures in a sequential manner
using appropriate precautions to ensure test and sample integrity and by segregating the activities
by time or space in conjunction with regulatory requirements. It is generally considered as a good
practice to have separate areas for:
Sample receipt and sample storage
Media preparation and Sterilization
Sample preparation
Aseptic operations
Incubation
Storage of reference cultures/Reference materials
Decontamination and washing
Sterility testing
Animal House

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3.5
Laboratories, shall take all precautions to avoid cross contamination. Dedicated pipettes, tips,
centrifuges, tubes etc should be located in each work area.
3.6
Laboratory located in facilities where Products or ingredients are manufactured shall not conduct
test for pathogens unless the laboratory is physically separated with limited access, equipped with
bio-safety cabinets and is supervised by a qualified microbiologist.
For procedures that involve the handling of pathogens and reference stock cultures, they shall be
operated within a safety cabinet of a class commensurate with the risk level of the microorganism
handled. Most of the microbes encountered in a non-clinical testing laboratory belong to Risk
Group 2 microorganisms. When working with samples containing microorganisms transmissible
by the respiratory route or when the work produces a significant risk from aerosol production, a
biological safety cabinet of Class II shall be used.
3.6.1 Properly maintained biological safety cabinet and other appropriate personal protective
equipment shall be used whenever:
(i)
Any infectious aerosol is likely to be produced while performing the test. These
include pipetting, centrifuging, grinding, blending, mixing, sonicating or opening
and transferring vials containing infectious materials, inoculating animals, and
harvesting infected tissues from animals or eggs, etc.
(ii)
Concentrated infectious agents such as enriched cultures of pathogenic

microorganisms, infected tissues etc. used in a test methodology.
3.6.2 Protective lab coat, gowns designated for lab use must be worn while working in the lab.
Protective clothing should be removed before leaving for non-laboratory areas. Eye and
face protection (goggles, mask etc.) should be used for anticipated splashes when
microorganisms handled outside BSC. Gloves must be worn to protect hands from
exposure to hazardous materials. Gloves shall be disposed off with other contaminated
waste after use. Disposable glove shall not be reused. Eye and face protection should
be used in rooms containing infected animals
3.7
Laboratories shall have an appropriate environmental monitoring programme with respect to the
type of tests being carried out. Records shall be maintained for it. Based on trends/anomalies
observed during the monitoring programme corrective action shall be taken and recorded. For
example, air borne/surface contamination can be monitored through exposure plates, air sampler
and surface swabbing etc.

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Acceptable backgrounds should be assigned and there shall be a documented procedure for
dealing with situations in which these limits are exceeded. Records of such situations, evaluation
of the effects, if any, on the test results and corrective actions taken shall be maintained.
Environmental contamination by microorganisms can be controlled by appropriate air- filters and
air-exchange systems and supervision by a qualified microbiologist.
3.8
The laboratory shall have pest control programme /schedule.
3.9
Additional Requirements for GMO Testing Laboratories

There shall be effective separation of the PCR testing area from neighboring laboratory areas to
minimize the spread of contamination from nucleic acids and or nucleases (both DNase and
RNase). Even minor degrees of cross contamination may result in erroneous results by nucleic
acid amplification. A separate room shall be used for PCR testing in a laboratory. Prevention of all
type of contamination is very essential. Procedure and precaution taken in avoiding the cross
contamination shall be documented. Such procedures shall include washing of lab ware,
generation of distilled, deionized or reagent water, decontamination of equipment between
samples during PCR analysis, cleaning of work surfaces and other relevant activities.
To avoid contamination, the laboratory should be organized to ensure unidirectional transfer of
samples. To fulfill this criterion, separate laboratories (or at least separate chambers) must be
made for each phase of the detection process, including sample storage, sample homogenization,
isolation of DNA, PCR reaction set-up, addition of isolated DNA, room for PCR instruments and a
room for post-PCR analysis (gel electrophoresis). Preventive actions such as decontamination
with UV radiation, changing laboratory clothing and gloves, using separate laboratory ware,
reaction reagents, pipette sets, etc., for each laboratory, ensure that no contamination is
transferred between different stages of the detection procedure.
The environmental conditions must also enable correct performance of the tests. Some parts of
the procedures are temperature-sensitive, especially where small volumes have to be measured.
Variation in the environmental temperature can cause large differences in pipetting accuracy,
causing differences in final concentrations of compounds in PCR. Thus, maintenance and
recording of constant temperature is essential
3.9.1
Reagents, consumables and equipment shall be located at appropriate designated areas

to serve their specific purposes. Nucleic acid samples should be kept in designated
refrigerated compartments after the sample preparation. They shall not be kept at areas

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where activity such as gel electrophoresis or PCR work is conducted. The movement of
nucleic acid samples or specimens should be as far as possible be unidirectional i.e. from
pre-amplification to post-amplification areas. Physically separate areas shall be provided
for the following:
a. GMO negative control
b. GMO Positive control/plasmid/vector
c. Sample extracts and
d. Kits, master matrix, taq polymerase, primers, probes, reagents
3.10
General Requirements for Toxicology Laboratories

Laboratories carrying out toxicological testing shall have an animal house segregated from other
activities of testing. The space shall be adequate for proper segregation of animals. Animal house
should be air-conditioned. The feeding and breeding is to be done by properly trained personnel
in animal care under the supervision of a veterinary doctor. A veterinary doctor shall periodically
check up the animals. The animals required for pyrogen test for drugs should be supplied as per
the test norms of national pharmacopoeia. The whole environment of animal house should always
be kept hygienic. Log books and other records shall be kept for the animal house maintenance
and animal care.
3.10.1 Facilities for Test Systems
Toxicological testing constitute an important part of biological testing for determining the nonclinical safety of a wide variety of industrial, environmental, pharmaceutical, agricultural and
consumer products. Biological test systems like laboratory animals, plants, fish and insects, etc
for in-vivo testing and cell cultures, micro organisms etc for in-vitro testing are often used for
toxicological testing. The laboratories should address the following issues to provide adequate
and appropriate accommodation and environment for housing, maintenance and assure their
health/viability and suitability through out the period of study.
3.10.2 Test System/Animal Facilities
3.10.2.1 A well planned, designed, constructed and maintained animal facility is required for efficient,
economical and safe operation in animal management for biological research and testing.
The size of animal facility depends on the scope of institutional activity and the types of

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animals housed. Animal facility should be physically separated from personnel and
laboratory areas. Construction material for interior surfaces (durable, moisture proof, fireresistant, seamless) should facilitate efficient and hygienic operation. Functional areas for
animal housing, quarantine, sanitation, storage of feed and materials, etc. should be
clearly designated. Corridors should be wide enough for free movement of personnel and
equipment. Corridors for clean and dirty operations should be clearly marked with
arrangement to prevent intermixing and contaminations during cleaning, disinfection,
change of cages and racks, autoclaving, waste removal and transport of animals, etc.
3.10.2.2
There should be proper separation of test animals by studies and species, separation of
healthy stock from diseased, separation of high hazard material and bio-hazardous
materials that can contaminate and affect the integrity of test system or invalidate the test
data.
3.10.2.3
Space allocation for animals shall, at the minimum, allow every individual to turn around
and express normal postural adjustments, ready access to food and water, enough clean
bedded or unobstructed area to move, stand and rest in.
3.10.2.4
Environmental temperature and humidity conditions of animal house should be

complimentary to the normal variations of animal body temperatures for their well being
and suitability for the studies. For most of commonly used laboratory animals in biological
testing a temperature range of 223C and relative humidity of 30-70% is considered
appropriate.
3.10.2.5
Adequate ventilation, necessary for adequate supply of oxygen, is generally considered

satisfactory with 10-15 fresh air changes per hour in the secondary enclosure (animal
room) subject to frequent cleaning of bedding and cages, control of recycled air, and use
of air filtration devices. Caging with forced ventilation that uses filtered room air and other
types of special primary enclosures with independent air-supplies can effectively address
the ventilation requirements of animals without ventilating secondary enclosures.
3.10.2.6
Illumination should generally be diffused throughout the animal holding area. Lighting in
animal rooms should provide for adequate vision and neuro-endocrine regulation of
diurnal and circadian cycles of animals. Most of the commonly used laboratory animals
are nocturnal. Photoperiod is a critical regulator of reproductive behaviour in many
species of animals and can affect food intake and body weight gain. A 12 hrs dark-light

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cycle is generally acceptable. A time-controlled lighting system should be used to ensure

regular diurnal cycle and the timer performance should be periodically checked and
calibrated for accuracy. Light levels of about 325 lux (30 ft. candles) in an occupied room
or 400 lux (37 ft candles) in an empty room, about one meter (3.3 ft) above the floor,
appear to be sufficient for animal care and do not cause clinical signs of phototoxic
retinopathy in albino rats (most susceptible species).
3.10.2.7 Noise, produced by animals and animal care related activities, is inherent in the operation
of an animal facility but it should be regulated to the minimum practicable. Measures such
as separation of human and animal areas, separation of noisy animals (dogs, swine,
goats, and non-human primates) from quieter animals (rodents), environmental design to
absorb noise, minimizing personnel and frequency of visits to animal rooms, etc. should
be adopted to keep the noise below 85dB.
3.10.2.8 Animals should be fed palatable, non-contaminated and nutritionally adequate food daily
or according to their particular requirements unless required otherwise by specific
protocol. All feed should be free from chemical, microbial, fungal contaminants and
natural toxins. Purchase, transport, storage and handling of all food should minimize
introduction of contaminants and diseases, parasites, potential disease vectors, etc.
Autoclavable diets may require adjustments in nutrient concentrations degraded during
sterilization. The date of sterilization should be recorded and the diet used quickly.
Feeders should be designed and placed to allow easy access to food and to minimize
contamination with urine and feces. Feeders should also be cleaned and sanitized
regularly.
3.10.2.9 Animals should be provided potable, uncontaminated drinking water according to their
requirement. Periodic monitoring of drinking water for pH, hardness, microbial and
chemical contaminants should be done to ensure that water quality is acceptable. Water
can be treated or purified to eliminate contaminants if the protocol requires highly purified
water however, such process like chlorination, should be considered for adverse effects
on the test system and study results. Watering devices should be checked daily for their
proper maintenance, cleanliness and operation.
3.10.2.10 Appropriate and sufficient bedding should be provided in animal housing to keep the
animals clean and dry between cage/bedding changes. Bedding should be changed as
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frequently as necessary due to leakage of water bottles, diarrhea, etc. Bedding should be
transported and stored off the floor on pellets, racks or carts to minimize contamination. It
should be autoclaved and dried (to evaporate moisture) before use.
3.10.2.11 Adequate sanitation of animal house is necessary for good health of test system and
integrity of studies. Sanitation includes bedding change, cleaning (removal of dirt and
debris) and disinfection (reduction/elimination of unacceptable concentration of microorganisms). The frequency of sanitation operation is dependent on the type of housing,
species of animal, type of bedding material, environmental conditions, etc. Deodorants
should not be used to substitute good sanitation and adequate ventilation. Cleaning and
disinfection of pens, cages and other primary enclosures should be done with frequent
flushing with water and periodic use of detergents and disinfectants. The frequency of
cleaning cages, racks, and associated equipment like feeders, water bottles, etc should
be preferably once a day. Acid wash may be necessary to clean the cages of rabbits,
guinea pigs and hamsters that produce urine with high concentration of proteins and
minerals.
Disinfection can be achieved with chemicals, hot water or a combination of the two.
Washing time should be sufficient to kill vegetative forms of common bacteria and other
organisms that are presumed to be controlled by the sanitation program. Disinfectants
should be thoroughly rinsed before reuse of equipment. Details of disinfectant use should
be recorded. All areas of animal facility should be routinely cleaned and disinfected.
Cleaning utensils should be assigned to specific area to limit possibility of cross
contamination and be cleaned or replaced themselves to be in good working condition.
Effectiveness of sanitation can be monitored by visual inspection, odors and
microbiological testing.
3.10.2.12 Programs designed to prevent, control or eliminate pest infestation should be
implemented in an animal facility. Non-toxic means of pest control such as insect growth
regulators, non-toxic substances (amorphous silica gel) traps, etc. should be used. Use of
pesticides should be avoided but if necessary, it should be documented and any impact
on study animals is to be evaluated.

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3.10.2.13 Institutional arrangements for emergency, weekend and holiday care of animals should be
prominently displayed along with responsible persons names, phone etc. to contact in
such event.
3.10.2.14
Suitable rooms or areas should be available for the diagnosis, treatment and control of
diseases, in order to ensure that there is no unacceptable degree of deterioration of test
systems.
3.10.2.15
There should be separate areas designated for treatment and other test related
procedures on the animals. Area for necropsy should be separate from animal housing
because killing animals in presence of other animals is considered stressful and
unethical. Necropsy areas should be equipped for euthanasia.
3.10.2.16
Other test systems including tissues, cells, sub-cellular preparations, micro-organisms,

etc. should be provided appropriate accommodation and environmental conditions so as
to preserve their identity & characteristics, and avoid all forms of contaminations that may
influence their integrity and/or results. This may require appropriate equipment for their
transport, storage and handling, specific procedures for characterization, labeling and
handling, periodic sampling to assure their integrity, etc. Test systems requiring cryopreservation
may
be
kept
in
multiple
samples/aliquots
in
small
vials/tubes/receptacles/containers. Since the labels on such small containers may not

allow much detail, codes may be used on the containers and code-wise-records of test
system details (identity, characteristics, source, date of receipt, storage condition, expiry,
etc.), usage, reculture and disposal should be maintained.
3.10.2.17
There should be storage rooms or areas as needed for supplies and equipment. Storage
rooms or areas should be separated from rooms or areas housing the test systems and
should provide adequate protection against infestation, contamination, and/or
deterioration.
3.10.2.18
There should be appropriate and adequate arrangement for un-interrupted power supply
to the animal/test system facility. The electric system should be safe with back-up power
supply in case of power failure through normal channel.

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4.
Test Methods and Method Validation (ISO/IEC 17025 clause 5.4)

Accreditation is granted for internationally or nationally accepted standard test procedures or nonstandard procedures /laboratory developed methods that have been appropriately validated and
which are performed regularly.
4.1
Standard Methods
The standard methods shall be used wherever available in order to ensure inter lab reproducibility
of the results. The laboratory shall maintain current versions of the controlled standard methods.
The laboratory must verify that it can properly operate the method, and can demonstrate (where
specified) the limits of detection, selectivity, repeatability and reproducibility. Laboratories shall
pay attention to the limitations, concentrations range and sample matrix specified in the test
standards. The laboratory shall validate standard methods applied to the matrices not specified
therein.
4.2
Lab developed/non standard methods

These include but not restricted to:
4.3
Methods prescribed by a customer (to be specified )
Modified standard test methods (to be specified )
Methods from scientific publications, but which have not been validated.(to be specified )
Kits
The use of commercial test systems (kits) will require further validation if the laboratory is unable
to source the validation data. When the manufacturer of the test kits supplies validation data, the
laboratory will only perform secondary validation (verification).
Laboratories shall retain validation data on commercial test systems (kits) used in the laboratory.
These validation data may be obtained through collaborative testing, from the manufacturers and
subjected to third party evaluation (e.g. AOAC. Refer www.aoac.org for information on methods
validation). If the validation data is not available or not applicable, the laboratory shall be
responsible for completing the primary validation of the method.

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It has been found in some cases (e.g. veterinary microbiological testing) that a specific test kit
performs differently under local environmental conditions, to that of the original environmental
conditions when subjected to primary validation. In such cases the laboratory should conduct the
validation to prove that the kit performs under local environmental conditions.
4.4
Validation of Microbiological Test Methods (ISO/IEC 17025 Clause 5.4.3)

4.4.1
The validation of microbiological test methods should reflect actual test conditions. This
may be achieved by using naturally contaminated Products or Products spiked with a
predetermined level of contaminating organisms. The analyst should be aware that the
addition of contaminating organisms to a matrix only mimics in a superficial way the
presence of the naturally occurring contaminants. However, it is often the best and only
solution available. The extent of validation necessary will depend on the method and the
application. The laboratory shall validate standard methods applied to matrices not
specified in the standard procedure.
4.4.2
Qualitative microbiological test methods, confirmation and identification procedures

should be validated by determining specificity, relative trueness, positive deviation,
negative deviation, limit of detection, matrix effect, repeatability and reproducibility, if
appropriate. (See Appendix -B for definitions).
4.4.3
For quantitative microbiological test methods, the specificity, sensitivity, relative trueness,
positive deviation, negative deviation, repeatability, reproducibility and the limit of
determination within a defined variability should be considered and, if necessary,
quantitatively determined in assays. The differences due to the matrices must be taken
into account when testing different types of samples. The results should be evaluated
with appropriate statistical methods.
4.4.4
If a modified version of a method is required to meet the same specification as the

original method, then comparisons should be carried out using replicates to ensure that
this is the case. Experimental design and analysis of results must be statistically valid.
Even when validation is complete, the user will still need to verify on a regular basis that
the documented performance can be met, e.g. by the use of spiked samples or by
incorporating reference materials in relevant matrices.
Appendix- C provides some guidelines for method validation in microbiology

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4.5
Test Methods and Method Validation of GMO testing

The current GMO testing using PCR technology covers several types of analysis including inter
alia, qualitative, semi quantitative and real time quantitative test. Requirements for method
validation for different analysis vary slightly.
4.5.1 Test Methods:

4.5.1.1
Laboratories should be clear about which matrices can and can not be tested. For example,
it is generally accepted that refined oils can not be tested due to the absence of DNA and
should document that such tests should normally be refused.
4.5.1.2
There are some processed food matrices (e.g. soy sauce) where the integrity of the DNA
needs to be assessed to decide whether the test has any validity.
4.5.1.3
GM testing methods should include background information on GM and the traits being
tested for. The laboratory shall maintain background information on which GM materials
(crops) are on the market, so that inappropriate testing is not undertaken, or inappropriate
claims not made from results.
4.5.1.4
When a GM screening test is used as a preliminary detection tool, the use of such test needs
to be validated to demonstrate that it would detect a defined range of foreign DNA. Individual
detection limits should be determined as the detection limits may vary in such screening test.
4.5.1.5
If a GM screening test is negative and no further testing conducted, the result should be
reported as no foreign DNA sequence detected with respect to the specific test conducted,
with a specification of which traits have been excluded.
4.5.1.6
If a GM screening test is positive, then the laboratory should proceed to determine the
specific trait present and can also specify the range of traits tested.
4.5.2
Validation of Methods:
4.5.2.1
The laboratory should be clear about which matrices are suitable for quantification. Basing
quantification on a line from reference materials prepared from one matrix may not be
appropriate for the same trait in a different (e.g. processed) matrix.
4.5.2.2
As the availability of GM reference materials for quantification will always lag behind the traits
that are on the market, the laboratory may mix its own quantification standards from 100%

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GM material, provided that the purity of the materials (GM and non GM) shall be established
and proper validation undertaken.
4.5.2.3
Commercial test systems (kits) may not require further verification if validation data based on
collaborative testing are available. Otherwise, laboratory shall be responsible for validation of
the method. The laboratory shall demonstrate their capability to achieve the limit of detection
quoted by the manufacturer or the laboratory has to establish its own limit of detection to
minimize false positive and false negative results.
4.5.2.4
Laboratories shall determine the method performance characteristics such as limit of

detection, precision, etc for quantitative tests.
4.5.2.5
DNA assessment for analysis of items containing several ingredients or having been
processed (e.g. food), laboratories shall verify that the extraction and clean up procedures
used are capable of extracting good quality amplifiable DNA and the resultant extracts are
free from inhibiting substances. Procedures and methods used shall be so designed as to
minimize the risk of false negative results due to the presence of inhibitors of nucleic acid
amplification or restriction enzyme activity. Extraction method shall be validated for their
ability to remove inhibiting substances.
4.5.2.6
Quality of the extracted DNA from all samples shall be assessed by some well-established
method (gel based assessment and amplification of a house keeping gene are common).
This provides a means of assessing whether the DNA has lost integrity, and in such
situations further testing would be inappropriate. A laboratory may have established an
extraction method for a single sample, and then assume that for all such samples the
extraction is effective and DNA is suitable for analysis.
4.5.2.7
For extraction method that has not been shown to remove consistently the inhibitors, an
inhibitor control shall be used. The inhibition can be estimated by the amplification of another
target nucleic acid expected to be present in all samples or a known DNA spiked in test
samples at known concentrations.
4.6
Test Methods for Toxicological Testing
4.6.1
Toxicological laboratories should use standard study protocols and standard operating
procedures/test methods referred in the BIS test procedures, OECD Test guidelines, Schedule-Y,
Goitonde Committee report etc. Modifications to such standard guidelines should be described

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and justified with validation. Details of test method validation should be retained with the raw data
wherever applicable.
4.6.2
Laboratories should maintain details of experimental design including justification for selection of
test system and its characteristics (species, strain, sub strain, source, sex, age, weight, etc),
justification for the method, frequency and dose of exposure, chronology of events, methods and
materials, type and frequency of analysis/measurements and statistical evaluation etc.
4.7
Uncertainty of Measurement (ISO/IEC 17025 Clause 5.4.6)

It is important for testing laboratories to understand the concept of uncertainty of measurement.
The laboratory management should be aware of the effect that their own uncertainty of
measurement will have on test results produced in their laboratory.
It is well recognized that the current state of knowledge regarding uncertainty of measurement
across the full range of biological discipline is variable and a consensus agreement on the
definitive methodology to be used for estimating uncertainty is still some way off. All concerned
are encouraged to familiarize themselves with current developments through all available
sources such as relevant guidelines, specifications, publications, scientific texts or journals etc.
4.7.1
The following details the current requirements for laboratories accredited under Biological testing
program:
4.7.1.1 The laboratories need to make a formal estimate of measurement uncertainty for all tests
in the scope of accreditation that provide numerical results. Where the test results are not
numerical or are not based on numerical data e.g. detected/not detected, pass/fail,
positive/negative or based upon visual, tactile or other qualitative examination uncertainty
estimation is not required.
Nevertheless, individual sources of variability, e.g. consistency in the performance of
media/reagents, and analyst interpretations should be identified and demonstrated to be
under control. The laboratories should also be aware of the incidence of false positive
and false negative results associated with the qualitative test they perform.
4.7.1.2 Where the laboratory needs to estimate the measurement uncertainty, it is required to
document the procedures and processes on how this is to be done. There are various
published approaches to estimate the uncertainty in testing. ISO/IEC 17025 2005 does

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not specify any particular approach. All approaches that give a reasonable estimation of
uncertainty are considered valid. What is important is that laboratories document with
reference to published approaches, what their approach to estimating uncertainty in
measurement will be. The suitability and rigor of the adopted approach will be assessed
accordingly.
Out of various approaches available in the literature appendix- F sets out a possible
approach suggested to be consistent with the approaches available internationally. This
approach is not mandatory but in case any alternative approach is adopted the same will
be considered as an equally valid if sourced from published guide lines and expected to
address the principles embodied within it.
4.7.1.3 Once the approach is adopted and a procedure is established the laboratory needs to
develop and commence implementation of a program for applying this procedure to all
relevant tests within the scope of accreditation.
4.7.2
Reporting of measurement uncertainty

Clause 5.10.3.1(c) of ISO/IEC 17025 2005 requires reporting of measurement uncertainty when it
is required for the correct application or interpretation of test results. One such instance is where
test results are used to determine if a sample conforms to a required numerical specification, and
where the specification limit falls within the limits of measurement uncertainty associated with the
test results obtained.
Biological testing laboratories are not required to report their measurement uncertainty on test
reports as a matter of routine
4.7.3
Measurement Uncertainty in calibrations:

Clause 5.4.6.1 of ISO/IEC 17025:2005 requires that the testing laboratories, which perform their
own calibrations, shall have and apply a procedure to estimate the uncertainty of measurement in
all calibrations. The full rigor of this requirement is expected to be applied where the equipment
item being calibrated has performance requirements that are critical to the accuracy or proper
performance of the test and are approaching the performance specification of that equipment
item. The example includes the calibration of analytical balances, incubators and thermometers
requiring high level of accuracy. For all these calibrations, a full measurement uncertainty budget

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is expected to be estimated. This would normally be expected to be estimated in accordance

with the Guide to the expression of uncertainty of measurement. (ISO 1995)
Uncertainty of measurement estimations for periodic checks conducted in-house on
calibrated equipments, are not required

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Equipment - Maintenance, Calibration and Performance (ISO/IEC 17025 Clause 5.5)

As part of its quality system, all accredited laboratories are required to maintain a documented
programme for the maintenance, calibration and performance verification of its equipment
necessary to carry out the tests included in the scope of accreditation.
5.1
Maintenance
5.1.1
Maintenance of essential equipments used in the laboratory shall be carried out at specified
intervals as determined by factors such as the rate of use. Detailed records shall be kept.
If a test method or operating environment requires a more stringent calibration/verification interval
than that set by the laboratory, more frequent calibration will apply.
5.1.2
Laboratory should have established procedures & schedule (cleaning & sanitization) to ensure
avoidance of cross-contamination arising from the equipments used to perform the tests..
5.1.3
Apparatus, including validated computerized systems, used for the generation, storage and
retrieval of data, and for controlling environmental factors relevant to the toxicological test should
be suitably located, and of appropriate design and adequate capacity.
5.2
Calibration and Performance Verification

Commonly used equipment for biological tests that requires calibration and/or performance
verification include balances, thermometers, pH meter, timer, ovens, incubators, autoclaves,
water bath, Laminar Flow chamber, Biosafety cabinets, thermocycler and volumetric glassware.
5.2.1
Autoclave
5.2.1.1 Autoclave shall not be used to sterilize clean equipment and to decontaminate used
equipment during the same sterilization cycle. Ideally the laboratories should have
separate autoclave for these two purposes. Records of autoclave operations including
temperature and time shall be maintained. Acceptance and rejection criteria for operation
conditions shall be set and implemented.
5.2.1.2 Pressure measurements alone can not guarantee that appropriate temperature has been
attained through the sterilization cycle. Measurement of temperature is essential for each
autoclave cycle to ensure that the unit has been correctly vented. Autoclaves therefore
need to incorporate a temperature recording device.

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5.2.1.3 Temperature controllers, temperature recording device and thermocouples need to be

calibrated initially and every six months using a reference thermometer or thermocouple,
which in turn an accredited calibration laboratory, has calibrated. The temperature
calibration results will reveal the pressure gauge deficiencies.
5.2.1.4 In addition to monitoring the temperature, the effectiveness of sterilization can be checked
with biological and chemical indicators. Temperature sensitive tape or indicator strips
shall be applied for each load. However they are used only to show that the load has
been processed but not as a monitor of the actual process applied.
5.2.1.5 Validation of autoclaves enables laboratories to demonstrate acceptable and consistent
temperature of sterilization. The main thrust of the need to validate autoclaves is to
ensure that the media used for microbiological analysis are not being over cooked in the
autoclaves. In particular the temperatures should not exceed 121C and that media are
not exposed to a high temperature for too long a time. Sufficient heat is needed to kill all
spores whilst protecting the media from excessive heat input thereby overcooking.
5.2.1.6 Performance of autoclaves shall be checked periodically with biological indicators.
5.2.2
Incubators, Water Bath, Hot Air Ovens

5.2.2.1 The stability of temperature, uniformity of temperature distribution and time required to
achieve equilibrium conditions in incubators, water baths ovens and temperature
controlled rooms shall be established initially and documented, in particular with respect
to typical uses. (for example position, space between and height of ,stacks of Petri
dishes). Temperature of incubators shall be verified against the specifications of the test
standards and checks on the shelves shall be recorded. Temperatures at different levels
and different positions at same level inside the incubator shall be verified at defined time
intervals and at least annually against the temperature specifications of the tests.
5.2.2.2 Performance of ovens shall be checked periodically with biological indicators.
5.2.3
Temperature Monitoring Devices

Where the accuracy of the temperature measurement has a direct effect on the result of the
analysis, the temperature measuring devices used in incubators and autoclaves shall be of
appropriate quality to achieve the specifications in the test methods. The graduation of the device
shall be appropriate for the required accuracy. Traceability of the temperature measurement

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device has to be established and overall uncertainty of measurement shall be estimated and must
be appropriate for the measurement.
5.2.4
Refrigerator, Freezer or Cold Storage Room

Permissible ranges of operation shall be specified and records of temperature checks shall be
maintained.
5.2.5
Weights and Balances

Weights and balances shall be calibrated traceably at regular intervals according to their intended
use.
5.2.6
Volumetric Equipment
5.2.6.1 Volumetric equipment such as automatic dispensers, dispenser /diluters, mechanical
hand pipettes and disposable pipettes may all be used in the biological laboratory.
Laboratories should carry out initial verification of volumetric equipment and then make
regular checks to ensure that the equipment is performing within the required
specification. Verification should not be necessary for glassware, which has been certified
to a specific tolerance. Equipment should be checked for the accuracy of the delivered
volume against the set volume (for several different settings in the case of variable
volume instruments) and the precision of the repeat deliveries should be measured.
5.2.6.2 For single-use disposable volumetric equipment, laboratories should obtain supplies
from companies with a recognized and relevant quality system. After initial validation of
the suitability of the equipment, it is recommended that random checks on accuracy are
carried out. If the supplier does not have a recognized quality system, laboratories should
check each batch of equipment for suitability.
5.2.7
Laminar Flow Hoods

It is important that laminar flow hoods are serviced annually. High Efficiency Particulate Air
(HEPA, 99.9%) filters shall be checked and cleaned or replaced as needed.
Airflow rate shall be monitored regularly or at-least annually with a calibrated velometer,
anemometer or other appropriate flow instrument, to ensure that the exhaust system
functions properly. Particle count shall also be checked on a routine basis to comply with
relevant standard.

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Cleanliness of hood surfaces shall be maintained before and after each use. They shall be
routinely monitored using appropriate method such as the use of Replicate Organisms Direct
Agar Contact (RODAC) plates or by surface swabbing method.
During operation the aerial microbial contamination shall also be checked using agar plates or
air sampler.
5.2.8
Appropriate disinfection shall be carried out before and after use,
Biohazard Cabinet
Biohazard cabinet shall be used for personnel protection when testing for hazardous
microorganisms. It shall be maintained monthly, quarterly, or annually depending on the class of
the cabinet. Parameters such as final filter and exhaust filter integrity, air velocity and uniformity,
air barrier containment, induced air leakage, UV radiation, light intensity and noise level shall be
monitored.
Biosafety levels are explained in Appendix-E
5.2.9
PCR Equipment
The performance of the PCR equipment such as thermocycler and the built in spectroscopic
components of PCR equipment shall be verified regularly.
5.3
Reference Materials and Reference Cultures
5.3.1
Reference Materials
5.3.1.1 Reference materials and certified reference materials, if required should be used to
provide essential traceability in measurements and are used, for example;
To demonstrate the accuracy of results,
To calibrate equipment
To monitor laboratory performance,
To validate methods and
To enable comparison of methods
If possible, reference materials should be used in appropriate matrices.

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5.3.1.2 Regardless of the source of the certified reference materials, care shall be exercised to
see that they are stored and handled to prevent deterioration. They shall be kept under
secure and appropriate storage conditions, and records shall be maintained of their
receipt and use.
5.3.1.3 Responsibility shall be documented for the maintenance of for certified reference
materials which typically include ordering or assisting in the ordering of new reference
materials, checking calculations of assays, keeping lists of laboratory-available certified
reference materials up to date, properly identifying reference material containers,
maintaining reference materials in their proper location, disposing old or outdated
reference materials, and so forth.
5.3.1.4 Reference materials usage records shall be maintained to ensure traceability. Each
analyst using a certified reference material shall enter the name of the reference material,
the date and time, issued and returned, with initials.
5.3.1.5 Analysts shall be instructed in the care of certified reference materials and procedures for
handling them.
5.3.1.6 Biological testing laboratories are expected to source their reference materials from the
following possible sources (generally in decreasing order of preference) where availability
permits:
a) Reference standards from national measurement institutes and from ISO Guide 34
accredited reference material producers:
b) Reputable chemical supply houses (particularly kit manufacturers and for pure
biochemical standards or reagents);
c) Customer supplied reference standards, preferably with certification;
d) In-house produced reference standards
5.3.1.7 Laboratories shall demonstrate traceability for certified reference materials obtained from
a recognized national institute.
5.3.1.8 DNA extracted from certified reference materials are stored to provide reference stocks.
Reference stocks shall be stored at a condition to minimize nucleic acid degradation.
Laboratories shall have a policy and procedures for purchase, handling, storage,
maintenance and use of certified reference materials and stocks.Reference stocks should
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be aliquoted to minimize damage due to freezing and thawing. Laboratories should verify
stability of stock DNA. Procedures for verification of stocks should be documented.
5.3.1.10 The following records shall be maintained:
a) The sources, lot numbers, dates of receipt and expiry, dates put in use, conditions
and integrity of packaging of certified reference material;
b) Preparation records of reference stocks with dates of preparation, expiration, and
name of operator;
c)
Verification records of reference stocks; and
d)
Records of monitoring of environmental conditions for storage of reference stocks.
5.3.1.11 Positive DNA reference materials/plasmids/vectors shall be verified by checking with at

least one reference material from a different manufacturer or source, if available, before
use. The requirements as in 5.3.1.10 should be fulfilled for maintaining the records.
5.3.2
Reference Cultures
5.3.2.1
Reference cultures are required for establishing acceptable performance of media

(including test kits), for validating methods and for assessing/evaluating on-going
performance. Traceability is necessary, for example, when establishing media performance
and method validations. To demonstrate traceability, laboratories must use reference
strains of microorganisms obtained directly from a recognized national or international
collection, where these exist.
5.3.2.2
Reference strains when
obtained shall preferably have information related to the
microorganism which include reference number (e.g. ATCC,MTCC etc.), passage number,
colony characteristics on recommended media, microscopic features by staining technique,
phenotypic characteristics relevant to the scope of testing, pathogenic status of the strain &
any specific requirement for culture handling, storage conditions and relevant applications
(such as quality control, vitamin assay, antibiotic assay etc.)
If required, MTCC can provide the information related to diagnostic characteristics of the
culture, any specific requirement for culture handling, storage conditions and relevant
applications. In addition the information about the risk group that a particular strain belongs
is also available with MTCC.
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Reference cultures obtained from a recognized national or international collection shall be

verified for their characteristics on receipt as per the details in the certificate provided by
the culture collection and/or as per the requirements of the test method or activity. For
example, reference strain of Salmonella used for method verification should provide results
as mentioned in standard test procedure including biochemical tests. In case of antibiotic
assay, sensitivity of reference strain to a particular antibiotic should be verified & confirmed.
5.3.2.3
Reference cultures can be obtained from the following sources

a)
American Type Culture Collection (ATCC)
b)
Microbial Type Culture Collections, (MTCC),( IMTECH) Chandigarh
c)
National Collection of Industrial Microorganisms (NCIM)- National Chemical

Laboratory, Pune
5.3.2.4
d)
Christian Medical College, Vellore
e)
National Institute of Communicable Diseases, Delhi
f)
Central Research Institute, Kasouli, Himachal Pradesh
g)
National Culture Type Collections, (NCTC), U.K.
h)
Any other ISO Guide-34 accredited Reference Materials Producers
Reference cultures shall be sub-cultured once to provide reference stocks. Reference

stocks shall be preserved by a technique such as freeze-drying, liquid nitrogen storage,
frozen beads storage etc., which maintains desired characteristics of the strains.
Laboratories shall have a policy and procedures for purchase, handling, storage,
preservation, maintenance and use of reference cultures and stocks.
Reference stocks shall be used to prepare working stocks for routine work. Bacterial
working stocks if sub cultured should be done only up to a defined number of generations
which is recommended up to five passages from the original reference culture. Laboratory
should take responsibility to verify that the working stock used in daily QC checks or other
bioassay will meet the requirements of the test method i.e. there is no change in
biochemical, serological activity of the strain and no change in cell morphology and colony
characteristics on growth/ selective media. Use of reference culture after 5 passages may
be extended if the same strain is found to retain all desired characteristics (morphological,

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biochemical and serological characteristics as required by the scope of activity) which is

verified by the laboratory using suitable tests and justification for extending its use is
documented for the defined period from the date of verification.
Procedures for verification of working stocks shall be documented.
5.3.2.5
Culture maintenance records:

Laboratories should maintain records of all their reference culture maintenance activities,
including certificates from the reference culture Collection, verification records, and subculturing records including any purity/verification checks. Unambiguous culture code should
be assigned to each organism for easy identification and traceability.
5.3.2.6 Reference cultures of microorganisms available not directly from, but claimed to be
traceable to a national collection may be used for quality control checks, but the
requirements on number of passages and the relevant verification procedures required as
mentioned in 5.3.2.4 shall also be observed. They shall not be further sub-cultured if
no
information on passage number is available from the supplier

5.3.2.7 Specific records mentioned below shall be maintained by the laboratory:
a) Reference culture master records containing information of the source, lot number,
reference number, laboratory code for a particular strain, dates of receipt and expiration
(if available), date of revival;
b) verification records of working stocks for the parameters tested and the result summary
with original observation or cross reference to laboratory note book containing raw
data;
c) history of subculture from reference stocks with dates of preparation and expiration,
media code, purity confirmation, and name of operator;
d) details of preservation of reference stocks and records of monitoring of environmental
conditions for storage of reference cultures, reference and working stocks.
General requirement on reference culture maintenance, subculture and storage is given as
Appendix-D

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5.4
Reagents and Culture Media
5.4.1
Laboratories should ensure that the quality of reagents used is appropriate for the test concerned.
They shall verify the suitability of each batch of reagents critical for the test, initially and during its
shelf life, using positive and negative control organisms, which are traceable to recognized
national or international culture collections.
5.4.2
Laboratories shall have a policy and procedure(s) for the selection and purchasing of services
and supplies. Quality and grade of reagents/media should be appropriate for the tests concerned.
They shall not contain any impurities that may inhibit bacterial growth. Guidance on precautions,
which should be observed in the preparation or use of reagents, should be documented.
The precautions related to toxicity, flammability, stability to heat, air and light, reactivity to other
chemicals, etc. should be taken while their handling and storage.
Persons responsible for preparation of reagents shall be identifiable from records.
5.4.3
The sources and history of consumables having an effect on the validity of tests such as media,
antisera, biochemical kits and membrane filters shall be recorded. A logbook shall be maintained
to record all such materials received at laboratories. This logbook shall include information such
as supplier, lot number, date received, date put in use, date of verification and date of expiration.
5.4.4
Media, supplements and additives

5.4.4.1 All dehydrated complete or pre-prepared media and purified agars shall be checked for
their physical states and verified for their microbiological performance prior to release for
use. Selective media shall be checked using positive strains with typical characteristics
and completely inhibited strains, where appropriate. Commercially pre-prepared media
should have evidence of evaluation of quantitative performance. All laboratory prepared
media starting from basic ingredients shall be checked for their recovery i.e. quantitative
performance. Criteria of recovery and records of verification shall be maintained.
Laboratories should establish and record an appropriate re-ordering schedule to prevent
the holding of stocks beyond their expiry dates.
Schedules for checking media for decomposition, discoloration, deterioration and caking
shall be documented. It is important to prevent dehydrated culture media from absorbing
moisture during storage. Dehydrated media should be stored in a dry, cool and dark
environment. Acceptance ranges of storage conditions and criteria for rejecting media

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should be documented. Records of monitoring the storage conditions and checks of

media shall be maintained.
5.4.4.2 All media recipes and procedures for preparation shall be fully documented and
authorized. Records shall be kept of all relevant details of each batch of medium
prepared. The records should include medium name, lot number, manufacturer,
ingredient quantities (if applicable), final pH (post sterilization), and sterilization process,
date of preparation and name of operator. Prepared media not put in use immediately
shall be labeled with medium names or codes, date of preparation, and date of expiration
if applicable. Information on the life expectancy of prepared media under specific storage
conditions shall be specified and documented. Guidance on the preparation, sterilization
of media and recommended storage times can be found in ISO 7218: 1996 Microbiology
of food and animal feeding stuffs General rules for microbiological examinations and
American Public Health Association Standard Methods for the Examination of Water and
Wastewater (APHA) section 9020 B.
5.4.4.3 Quality of water used for testing should be specified and checked regularly for
compliance against the specific requirements. such as conductivity, pH and microbial
load etc. Only water that has been tested and found to be free from bactericidal or
inhibitory compounds is to be used for the culture media, reagents and diluents.
Glassware washing procedures need to ensure that no toxic residue left from detergents,
disinfectants and reagents etc
5.4.4.4 Serological and biochemical kits shall be verified with positive and negative strains with
typical and negative characteristics, if applicable.
5.4.4.5 Chemicals and reagents involved from sample preparation down to PCR testing shall be
molecular biology grade or equivalent and free from contaminating nucleic acids or
nucleases (both DNase and RNase). Extraction buffer or solution (If prepared in-house)
has to be autoclaved prior to use. Any special precautions in preparation or use of the
reagents shall be documented. Stability of the reagents to heat, air, light and other
chemicals etc should be included, if it is applicable and relevant.
5.4.4.6 All taq polymerase /master mix/kits/primers and probes shall be checked and verified for
their performance using GM positive materials prior to release for their use. Verification
procedures, criteria for acceptance, shelf lives and special storage conditions shall be
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documented. Records shall be maintained for verification and monitoring of the storage
conditions.
5.4.4.7 Membrane filtration units shall be stainless steel, glass, or autoclavable plastic, not
scratched or corroded and shall not leak. Diameter and pore size of membrane filters,
and diameter and absorption capability of absorbent pads shall meet the requirements
specified in the test standards. They shall be confirmed of their sterility prior to release for
use.
5.4.4.8 Sterile metal or disposable plastic loops, wood applicator sticks, sterile swabs, spreaders
etc. should be used as inoculating equipment. The metal inoculating loops should be
made of alloys that do not interfere with any biochemical tests.
5.5
Handling and Use of Test Systems for Toxicological Laboratories
5.5.1
Laboratories using animals for biological testing should invariably have an institutional Animal
Ethics Committee to review and approve the use of animals for studies in accordance with the
CPCSEA (Committee for the Purpose of Control and Supervision of Experiments on Animals)
and other national/international guidelines on the Care and Use of Animals for Biological
Research and Testing.
5.5.2
Laboratories should routinely use applicable guidelines for the breeding, care, management,
housing and use of laboratory animals being available from BIS, CPCSEA, NRC-USA, etc. both
for ethical considerations as well as integrity of test data. The handling and breeding is to be
done by personnel properly trained in animal care. A veterinary doctor shall periodically check up
the animals. The animals required for pyrogen test for drugs should meet the test norms of
national pharmacopoeia. The whole environment of animal house should always be kept
hygienic. Log books and other records shall be kept for the animal house maintenance and
animal care.
5.5.3
Proper conditions should be established and maintained for the storage, housing, handling and
care of biological test systems, in order to ensure the quality of the data.
5.5.4
Newly received animal and plant test systems should be isolated until their health status has
been evaluated. If any unusual mortality or morbidity occurs, this lot should not be used in
studies and, when appropriate, should be humanely destroyed. At the experimental starting date
of a study, test systems should be free of any disease or condition that might interfere with the
purpose or conduct of the study. Test systems that become diseased or injured during the

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course of a study should be isolated and treated, if necessary to maintain the integrity of the
study. Any diagnosis and treatment of any disease before or during a study should be recorded.
5.5.5
Records of source, date of arrival, and arrival condition of test systems should be maintained.
5.5.6
Biological test systems should be acclimatized to the test environment for an adequate period
before the first administration/application of the test or reference item.
5.5.7
All information needed to properly identify the test systems should appear on their housing or
containers. Individual test systems that are to be removed from their housing or containers
during the conduct of the study should bear appropriate identification, wherever possible.
5.5.8
During use, housing or containers for test systems should be cleaned and sanitised at
appropriate intervals. Any material that comes into contact with the test system should be free of
contaminants at levels that would interfere with the study. Bedding for animals should be
changed as required by sound husbandry practice. Use of pest control agents should be
documented.
5.5.9
Standard operating procedures should be available for the test system with respect to the
following:
i)
Room preparation and environmental room conditions for the test system.
ii)
Procedures for receipt, transfer, proper placement, characterization, identification and care
of the test system.
iii)
Test system preparation, observations and examinations, before, during and at the
conclusion of the study.
iv) Handling of test system individuals found moribund or dead during the study.
v)
Collection, identification and handling of specimens including necropsy and histopathology.
5.5.10 Cellular and microbial test systems should be characterized to assure their identity, viability, and
proper responsiveness to standard reference molecules/conditions for appropriateness of use in
the biological tests. They should be sampled and handled in a manner to avoid contamination and
mix-up and also to prevent any hazard to personnel. Wherever required, such test systems
should be used in designated, restricted and sterile areas and all waste should be neutralized /
sterilized before disposal.

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5.5.11 Records including test item and reference item characterization, date of receipt, expiry date,
quantities received and used in studies should be maintained.
5.6
Handling and Characterization of Test and Reference Items
5.6.1
Handling, sampling, and storage procedures of the test/reference item should be identified in
order that the homogeneity and stability are assured to the degree possible and contamination or
mix-up are precluded.
5.6.2
Storage container(s) of the test/reference item should carry identification information, expiry date,
and specific storage instructions.
5.6.3
Each test and reference item should be appropriately identified (e.g., code, Chemical Abstracts
Service Registry Number [CAS number], name, biological parameters).
5.6.4
For each study, the identity, including batch number, purity, composition, concentrations, or other
characteristics to appropriately define each batch of the test or reference items should be known.
5.6.5
In cases where the test item is supplied by the sponsor, there should be a mechanism, developed
in co-operation between the sponsor and the test facility, to verify the identity of the test item
subject to the study.
5.6.6
The stability of test and reference items under storage and test conditions should be known for all
studies.
5.6.7
If the test item is administered or applied in a vehicle, the homogeneity, concentration and stability
of the test item in that vehicle should be determined. For test items used in field studies (e.g., tank
mixes) these may be determined through separate laboratory experiments.
5.6.8
A sample for analytical purposes from each batch of test item should be retained for all studies
except short-term studies.
5.6.9
All records of test/reference item characterization, expiry and quantities received and used should
be retained with the study data.
5.7
Test and Reference Items (including Negative and Positive Control Items) for In-Vitro
Toxicity Tests
5.7.1
In general, the specific requirements for receipt, handling, sampling, storage and characterization
for test and reference items that are used in studies utilizing in vitro test systems are same as

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applicable to in-vivo tests except that aseptic conditions need to be observed in their handling to
avoid microbial contamination of test systems.
5.7.2
For positive reference items the definition implies not to grade the response of the test system to
the test item, but rather to control the proper performance of the test system. For negative
(vehicle) and positive control items, it may or may not be necessary to determine concentration
and homogeneity, since it may be sufficient to provide evidence for the correct, expected
response of the test system to them.
5.7.3
The expiry date of such control items may also be extended by documented evaluation or
analysis. Such evaluation may consist of documented evidence that the response of the
respective test systems to these positive, negative and/or vehicle control items does not deviate
from the historical control values recorded in the test facility, which should furthermore be
comparable to published reference values.

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6.
Sampling (ISO/IEC 17025 Clause 5.7)
6.1
In many cases, testing laboratories are not responsible for primary sampling to obtain test items.
Where they are responsible, it is strongly recommended that this sampling be covered by quality
assurance norms and must comply to applicable requirements. Customers taking their own
samples should be made aware of proper storage, sampling and transportation facilities.
Customers should be informed if the sample received is too small for meaningful analysis.
6.2
Transport and storage should be under conditions that maintain the integrity of the sample (e.g.
chilled or frozen where appropriate). The conditions should be monitored and records kept.
Where appropriate, responsibility for transport, storage between sampling and arrival at the
testing laboratory shall be clearly documented. Testing of the samples should be performed as
soon as possible after sampling and should conform to relevant standards and/or
national/international regulations.
6.3
Laboratories shall document the sampling procedures for taking test portions from laboratory
samples and shall have measures to ensure that the test portion is as representative of the
sample as possible, and the composition of the sample would not be altered in a way that would
affect the concentration or identification of the organisms/ targeted DNA being determined. In
GMO testing, for cases of whole beans or grains, sample shall be sufficiently large to provide
meaningful statistical data at the limit of detection of the method. The processed foods, canned
and bottled products, etc. could be collected in sufficient numbers belonging to the same batch for
analysis. In case different batches are used, details should be recorded and retained for
reference.
6.4
Special sampling procedures should be established for special/non-routine samples and made
available to the samplers as well as laboratory personnel. A copy of such documented procedure
shall be maintained with the raw data and retained for future reference.
6.5
Sampling should only be performed by trained personnel. It should be carried out aseptically
using sterile equipment. Environmental conditions for instance air contamination and temperature
should be monitored and recorded at the sampling site. Time of sampling should also be
recorded. Sampling procedure can form part of the test methods and shall include procedures for
sterilization of sampling equipment and precautions in performing aseptic techniques.

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6.6
In the case of seed testing laboratories sampling is the key activity and the laboratory
management must appoint specific personnel to perform particular types of sampling and seed
testing.
6.7
Seed testing laboratories must be able to demonstrate that it has a system for the approval of lot
identification, licensing of the seed samplers including the approval and /or provision of sampler
training programmes, and arrangements for maintaining and distributing up-to-date lists of
licensed seed samplers.
6.8
Seed testing laboratories should have procedures and practices to monitor the uniformity of seed
lots and to refuse the sampling and testing where doubt exists concerning uniformity.

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7.
Sample Handling (ISO/IEC 17025 Clause 5.8)
7.1
Laboratories shall examine and record the conditions and appearance of samples upon receipt.
Where appropriate (e.g. environmental samples for quantitative results), the time of sampling
should also be recorded. Parameters to be checked include nature and characteristics of sample,
volume/amount of sample, storage temperature of sample on receipt, conditions of sample
container i.e. whether it has been sterilized before sampling, characteristics of the sampling
operation (sampling date and condition), etc. If there is insufficient sample or the sample is in poor
condition due to physical deterioration, incorrect temperature, torn packaging or deficient labeling,
laboratories should either refuse the sample or carry out the tests as instructed by the customers
and shall indicate the conditions on test reports.
7.2
Samples awaiting test shall be stored under suitable conditions to minimize any changes to any
microbial population present. Storage conditions and maximum holding times for different
samples shall be documented and shall fulfill the requirements of test standards. Where a sample
has to be held secure, the laboratory must have arrangements for storage and security that
protect the condition and integrity of the secured samples concerned.
7.3
Frequently, it is necessary to split or transfer samples for different testing parameters. The Subsampling procedure should be designed to take account of uneven distribution of analytes. It
should be performed as per national/international standards, where they exist, or by validated inhouse methods. It is essential that procedures are available for preventing spread of
contamination, delivery of samples including special transportation such as refrigeration and
exclusion of light, disposal and decontamination processes and unbroken chain of identification of
the sub-samples/samples shall be provided.
7.4.
In sampling, the documentation sent to the seed testing laboratory must contain the following
information;
a) Name / identification and signature of the sampler
b) Name and address of the customer/exporter
c) Date of sampling, method of sampling and number of samples drawn
d) Unambiguous and unique reference number(s) identifying the lot. This may be a lot reference
number or a sequence or sequences of label numbers.

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e) Lot weight
f) Number and type of containers.
g) Tests required
h) Details of any environmental or other conditions during sampling which may affect the
interpretation of the test results.
i)
7.5.
Any other available information requested by a customer.
There shall be a written procedure and defined period for the retention and disposal of the
samples in the laboratory. Samples should be stored until the test results are obtained, or longer if
required. Laboratory sample portions that are highly contaminated should be decontaminated
prior to being discarded. Seed sample retention must be for not less than one year after testing
has been completed.

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8.
Disposal of Contaminated Waste
8.1
Waste Disposal
Decontamination of waste and the ultimate disposal are closely interrelated. Most glassware,
equipment, clothing are decontaminated or autoclaved within the laboratory for reuse. Other
contaminated or infectious waste shall be disposed of by using the documented procedures in line
with relevant regional or national regulatory standards, as applicable.
Identification and separation of contaminated waste materials are generally covered under the
following:
8.1.1
Contaminated sharps (broken glass, knife, scalpel etc) - should be collected in puncture
proof
containers and incinerated. Contaminated materials decontaminated by autoclaving and

thereafter washed and reused generally done within the lab.
8.1.1.1 Contaminated material decontaminated by autoclaving and thereafter disposed
with/without incineration as per regulatory norms. Except sharps, all other infectious
materials should be transported for disposal after autoclaving in a biohazard identifiable/
colour coded, leak proof pack.
8.1.1.2 Non-contaminated (non-infectious) waste - to be discarded as general waste.
8.1.2
Autoclaved waste can be disposed off through off-site incineration facility, in licensed landfill sites,
effluent treatment plants subject to meeting local regulations. Laboratory shall be provided with
biohazard identifiable or color coded waste disposal containers strategically placed within the lab
(for e.g. decontamination area). Suitable provision should be made for collecting the waste safely
from different areas within the lab. Contaminated toxicological waste, microbiological cultures etc
shall be disposed off as early as practically feasible. Biological waste such as animal carcasses,
anatomical as well as other associated wastes used in toxicological and other biological tests
should be discarded by appropriate decontamination and disposal facilities.
8.1.3
Waste disposal records shall be maintained for wastes disposed through licensed contractors.

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9.
Assuring Quality of Test Results (ISO/IEC 17025 Clause 5.9)
9.1
Laboratories shall establish and implement quality control plans to ensure and demonstrate that
the measurement process is in-control and test results generated are accurate and reliable. The
plans shall include types of quality control checks, their frequency and acceptance criteria, and
actions to be taken when results will be outside the defined acceptance criteria.
9.2
Internal Quality Control
9.3
Internal quality control consists of all the procedures undertaken by a laboratory for the
continuous evaluation of its work. The main objective is to ensure the consistency of day-to-day
results and their conformity with defined criteria.
9.4
Programme of periodic checks is necessary to demonstrate that variability (i.e. between analysts
and between equipment or materials etc.) is under control. All tests included in the laboratorys
scope of accreditation need to be covered. This can be achieved by:
9.4.1. Sterility controls

Uninoculated samples shall be run at a minimum of once for every test run. Sterility controls are
used to detect the presence or absence of possible laboratory contamination.
9.4.2
Split samples (Duplicates) for quantitative tests

Duplicate analysis usually involves a replicate sample, sub sampled in the laboratory. This
practice measures precision of an analytical process. For analysis performed in spiked matrices
the method precision is documented and controlled based upon relative percent difference in
recovery for quantitative determinations and confirmation of positive response in qualitative
analysis. Analysis of split samples is normally expected to be conducted at a frequency of at least
once per parameter/matrix/analyst.
9.4.3
Confirmation/verification of presumptive positive samples

Positive and negative characteristic strains, if applicable, shall be tested concurrently with any
biochemical, serological and morphological tests/characteristics for confirmation of presumptive
microorganisms. The number or percentage of colonies stipulated in test standard required for
confirmation process shall be followed. Laboratories can also define the minimum number of
colonies for confirmation if such requirements are not specified.

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9.4.4
Matrix spiked sample

The sample is prepared by adding a predetermined quantity of stock solution of representative
analyte to an actual sample matrix prior to sample preparation for analysis. The method is used to
measure accuracy of the method for qualitative estimations. The criteria may be used to establish
the precision of test methods. Acceptance limit can be established by running spiked samples of
cell suspensions in duplicate or triplicate, using two or more operators. The criteria used to set
acceptance limit for precision (relative standard deviation or range) shall be based upon statistical
principles and is clearly presented for each test method.
9.5
The following can be practiced as a quality control measure in testing laboratories where PCR
technique is being used. Example - GMO testing labs.
9.5.1
In-process Control Check

The following controls shall be run at a minimum of once for every test run as shown below: 9.5.1.1 Extraction negative (or blank) control
The extraction buffer employed for DNA extraction shall be prepared from sterile water
and shall be autoclaved prior to use.
9.5.1.2 Negative PCR control by use of non-GM material (0% GM content) exactly in the same
manner as the samples.
9.5.1.3 Detection limit control
A sample of known GM content or CRM can be used to establish the detection limit
meeting the limit of detection of the method. In the absence of a GM CRM, the laboratory
can spike appropriate amount of DNA enabling to achieve the desired detection limit.
9.5.1.4 Positive PCR amplification control
Reference DNA or DNA extracted from a CRM or a known positive sample representative
of a gene sequence under study shall be incorporated to demonstrate the unique
performance of the PCR assay.
9.5.1.5 Replicate analyses
PCR test samples shall be analyzed in at least duplicate for quantitative, semi
quantitative and qualitative testing. Because duplicate extractions and PCR of the same
sample can give qualitatively different results, one positive, one negative. In situations

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where false positive results occur due to contamination, rules out false negative results.
This situation is most likely to occur in cases where the test is working at concentrations
close to the limit of detection and/or there is some degree of inhibition of PCR due to coextractives from the sample.
9.5.1.6 Number of primer sets
It is normally expected that test results are based on the results of at least two, different
GM-specific primer sets, each providing consistent result. The requirement of using at
least two primer sets may be relaxed provided that other options for confirming the
identity of an amplicon on a gel, e.g. restriction enzyme cutting to produce fragments of
the expected size, shall be established to confirm test results.
9.6
Proficiency Testing Programme
9.6.1
Proficiency testing is defined as the evaluation of participant performance against preestablished criteria by means of inter-laboratory comparisons (ISO/IEC 17043:2010) and is thus
a very important tool in a laboratorys quality control programme to demonstrate the validity and
comparability of results.
9.6.2
Proficiency testing programme shall be scheduled and implemented on a regular basis relevant to
their scope of accreditation. Preference should be given to proficiency testing schemes, which
use appropriate matrices.
9.6.3
Laboratories should use external quality assessment not only to assess laboratory bias but also to
check the validity of the whole quality system.
9.6.4
In accordance with the policy of the Asia Pacific Laboratory Accreditation Co-operation (APLAC),
to which NABL is a member of their Mutual Recognition Arrangement (MRA), it is NABL policy
that applicant/accredited biological testing laboratories shall:
a) Demonstrate their technical competence by the satisfactory participation in proficiency testing
activity where such activity is available and that:
b) The minimum amount of appropriate proficiency testing required per laboratory is one activity
prior to gaining accreditation.
c) Accredited laboratories shall prepare a plan for PT/ILC participation so as to cover major
groups/subgroups in the scope. The plan shall consider the issue of changes in staff,
methodology, instrumentation, scope and critical tests etc. The recommended plan for

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participation would be done such that portion of scope (groups) are considered twice in a year.
In addition to this laboratory should consider the requirements of regulators for PT
participation. PT participation plan shall be prepared as per NABL 163 .
For practical reasons if laboratory is not able to follow this plan, lab shall have sufficient basis
for non-compliance.
9.6.5
Laboratories are expected to select the proficiency testing activities according to the following
criteria (in a generally decreasing order of preference):
(a) Mandated programmes. In some areas of biological testing, participation in a particular
programme may be mandatory.
(b) International inter-laboratory comparison/PT programmes.
(c) National inter-laboratory comparison/PT programmes.
(d) Proficiency testing programmes operated in accordance with ISO/IEC 17043.
(e) Inter-laboratory comparison programmes involving several independent NABL accredited
laboratories for statistical inferences.
9.6.6 .If unsatisfactory results are obtained, laboratories shall be able to show that the problems are
promptly investigated and rectified, and satisfactory performance for the test/method in question
can be achieved afterward. All findings in connection with unsatisfactory performance shall be
recorded.
(For better understanding laboratories can refer NABL 162 and NABL 163)
9.6.7
The results from proficiency testing activities and their analysis will be reviewed in each NABL
assessment.

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10.
Reporting the Results (ISO/IEC 17025 Clause 5.10)
10.1
Test Records
An adequate test record system in accordance with the various clauses of ISO/IEC 17025, e.g.
4.13, 5.4.7 is essential. Most laboratories have developed forms (proforma sheets) for all their
routine testing. These are generally the preferred option as their use prompts the recording of all
the required information, maintains consistency and increases recording efficiency.
10.2
Test records in the form of workbooks/worksheets shall be controlled and authorized by

designated key technical person and lab should ensure the traceability of raw data to the final
report.
10.3
Test Reports
10.3.1 Clause 5.10 of ISO/IEC 17025:2005 standard sets out the requirements for test report issued by
testing laboratories.
10.3.2 Test reports must give the customer all relevant information and every effort should be made to
ensure that the test report is unambiguous. All information in a test report must be supported by
the records pertaining to the test. All information required to be reported by the test specification
must be included in the report.
10.3.3 It is important to note that in many instances the test standards, regulatory requirements and
industry accepted practice will determine the report format and content.
10.3.4 Laboratories must retain an exact copy of all reports issued. These copies must be retained
securely and be readily available for the time specified in the laboratorys documented policies.
10.3.6 Where an estimate of the uncertainty of the test result is expressed on the test report on demand,
any limitations (particularly if the estimate does not include the component contributed by the
distribution of microorganisms within the sample) have to be made clear to the customer.
10.3.7 Laboratories carrying out GMO testing activities with PCR shall accurately describe the primer
sets used and the results obtained. The specificity of the target sequence shall be reported, i.e.
35S promoter: detected, or Roundup Ready: not detected or Bt-176: not detected instead of a
general statement does not contain GMO. The latter wording would imply that primer sets
covering all potential GM events had been tested. Similarly, quantitative results shall be reported
as x.x % of Roundup Ready Soybean instead of x.x % GM material.
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10.3.8 When test results are below the reporting limits, an indication of the reporting limits shall be given
in test reports.
10.3.9 The sample preparation procedure should be given for the proper interpretation of test results in
GMO testing laboratorys test reports.
10.3.10 NABL symbol in the test reports shall be used in accordance with NABL 133.
10.4
Electronic Reporting
Traditionally, laboratories issued test reports in hard copy format with manual signatures. With
increased use of electronic media such as email and the Internet, and the use of electronic
databases, laboratories are now issuing the reports electronically. Such practices challenge the
generally accepted reporting criteria for accredited laboratories.
10.4.1 ISO/IEC 17025: 2005 clause 5.10.7 attempts in a general way to specify the specific requirements
for electronic reporting. While it is difficult to specify in detail a set of requirements to address
every eventuality (as laboratories will tend to develop electronic reporting systems to suit their
own circumstances and those of their customers), the following is intended to provide guidance on
common issues of concern.
10.4.1.1 Transmission of Report
It is the responsibility of the issuing laboratory to ensure that what was transmitted electronically is
what the customer received.
Email systems have proven to be robust in this regard, but laboratories need to consider whether
customers will have the appropriate software and version to open attachments without corruption.
Laboratories should verify (at least initially, and periodically thereafter is recommended) the
integrity of the electronic link e.g. by asking the customer to supply a copy of what was received
and comparing it with what was transmitted. It is also important that the laboratory and its
customer agree as to which parts of the electronic transfer system they are responsible for and
the laboratory must be able to demonstrate data integrity at the point the data comes under the
control of the customer.

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10.4.1.2 Security
Laboratories should avoid sending test reports in an electronic format that can be readily
amended by the recipient. Where possible, reports should be in a read only format e.g. pdf files.
Where this is not possible e.g. the customer may wish to transfer the reported results file into a
larger database, then laboratories are recommended to indicate these electronic reports have
an interim status and are followed-up by a hard copy (or more secure) final report.
Laboratories must retain an exact copy of the report that was sent to the customer. This may be
a hard copy (strongly recommended) or an electronic copy. These copies must be retained
securely and be readily available for the time specified in the laboratorys documented policies.
10.4.1.3 Electronic Signatures
The reports must not be released to the customer until authorized by individuals with the
authority to do so. For electronic reports there must be a clear audit trail with a positive
authorization record to demonstrate this is the case. Where this is managed through password
access levels in the laboratorys electronic system, appropriate procedures should be in place
to prevent abuse of password access.
The electronic report should show the identity of the individual releasing the report (authorized
signatory approved by NABL). This may involve an electronic signature. The security of these
signatures should be such as to prevent inadvertent use or misuse.

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APPENDIX- A
CLASSES OF TESTS IN BIOLOGICAL DISCIPLINE
The biological testing discipline is described in terms of classes (Groups) and subclasses (subgroups) of
test. Application for accreditation may be made for one or more classes of tests or for subclasses or
specific test within a single class or subclass.
Where the existing group does not appear to cover the needs of a laboratory NABL secretariat welcomes
proposals for additional classes or tests to be included in this discipline.
The scope of accreditation may be reviewed and extended on request, provided that the laboratory
complies with conditions for accreditation for the classes of test or specific tests involved.
I.
Food and Agricultural Products

Animal Feeds
Bakery & Confectionery Products
Beverages (Alcoholic / Non-Alcoholic)
Canned & Processed Foods
Cereals, Pulses & Cereal Products
Coffee & Cocoa Products
Edible Colours & Flavours
Edible Oils & Fats
Eggs & Egg Products
Essential Nutrients Including Vitamins
Fish & Sea Foods
Food Additives & Preservatives
Fruit & Fruit Products
Gelatin and Other Gums
Genetically Modified Foods and agricultural products
Herbs, Spices & Condiments
Honey & Honey Products

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Infant Foods
Jams, Juices, Sauces & Concentrates
Meat & Meat Products
Milk & Dairy Products
Natural Waxes
Nuts & Nut Products
Oil Seeds & By-Products
Pet Foods
Poultry & Poultry Products
Starch & Starch Products
Sugar & Sugar Products
Snacks and Instant Mixes
Tea
Tobacco & Tobacco Products
Vegetables & Vegetable Products
Other Specified Food Items
II.
Drugs and Pharmaceuticals

Antibiotics
Ayurvedic Drugs
Biotechnology derived pharmaceuticals
Chemotherapeutic Agents
Drug Intermediates & Raw Materials
Endotoxins
Enzymes
Filtrable Solutions & Soluble Preparations
Hormones
Herbal drugs

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Immunological Products
Microbial limit test
Natural Drugs
Non-Filterable Preparations Including Ointments
Preservative efficacy
Pyrogen tests
Sterility tests
Surgical Dressings & medical accessories
Synthetic Drugs
Homeopathic Drugs
Vaccines
Veterinary Drugs
Biopharmaceuticals
Vitamins
Bioassays of Other Products (Other Than Those Products Mentioned Above)
Other Specified Tests
III.
Water
Drinking water
Packaged Drinking Water
Packaged Natural Mineral Water
Water for Swimming Pool and Spas
Water for Construction Purpose
Water Purifiers
Ground Water/ Surface Water
Water for Medicinal Purposes
Distilled /Demineralised Water
Water for Processed Food Industry
Water for industrial purpose

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IV.
Environment and Pollution

Bio burden Estimation of Classified and non classified area
Air /surface
Effluents/Waste water
Solid waste
Sewage
Soil
V.
Biocides
Algicides
Bactericides
Fungicides
Herbicides
Insecticides
Sporicides
Viricides
Weedicides
Antiseptics,
Disinfectants
Sanitizers
VI.
Cosmetics and Essential Oils

Gram negative Pathogens
Microbial Count
Preservative Efficacy
VII.
Industrial Cultures
Dairy Starter Cultures
Starter cultures for Effluent Treatment Plant
Rhizobial Cultures

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Cultures for baking and brewing

Mushroom Spawn
Probiotics Cultures
Other specified cultures
VIII.
Seed Testing
Purity
Germination
GM Testing
IX.
Plants and Plant Materials

Identification of Bacterial /Fungal/Viral Pathogens
X.
Molecular Analysis
(Tests for Various Matrices)
Genotyping
GMO Testing
Promoter & Terminator Screening
Detection of adulterants
Pathogen Detection
Gene Expression /Gene Copy Number
Bacterial Mutagenicity Tests
Sister Chromatid Exchange Tests
Transformation Assays In Cell culture
XI.
Cell Culture
Purity
Cell permeability test

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XII.
Resistance to Microbial Attack

Textiles and Fabrics
Leather & Leather Products
Electrical Components
Timber and Allied Material
Packaging Materials
Paints and surface coatings
Other Specified Materials
XIII.
Biological Tests on Other Miscellaneous Test Items

Adhesives Glues and Sealant
Fuels and Oils
Lubricants
Pulp & Paper
Soaps & Detergents
Textiles & Fabrics
Wood & Wooden Products
Toys and Other Childrens Products
Packaging Materials
Paints and surface coatings
XIV
Biopesticides and Biofertilizers
XV.
Toxicology
Acute Toxicity (oral, dermal, inhalation)
Mucous membrane irritation test
Skin sensitization test
Eye irritation test
Environmental toxicity/Eco toxicology

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Fish Toxicity
Bird Toxicity
Daphnia Toxicity
Earthworm Toxicity
Mutagenicity
Ames test
Dominant Lethal Mutation test
Cytogenetics
Chromosomal aberration test
Micronucleus test
Sister chromatid exchange test
Cytotoxicity
XVI.
Viability
DNA estimation
Protein estimation
MTT Assay
Identification/Enumeration of Microbial Pathogens by

Test Kits/ Rapid tests
ELISA
Qualitative PCR
Quantitative Real Time-PCR
XVII.
Residue Analysis
Antibiotic residue analysis by micro assay
XVIII. Veterinary Testing

Specified tests in biochemistry, haematology, cytopathology, histopathology, serology,
parasitological, virology, immunology etc.
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XIX.
Nutraceuticals & Functional Foods

a) Probiotics
XX.
Nutritional Supplements

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APPENDIX B
GLOSSARY OF TERMS
Calibration
Set of operations that establish, under specified conditions, the relationship between values of quantities
indicated by a measuring instrument or measuring system, or values represented by a material measure
or a reference material, and the corresponding values realized by standards
NOTES
1
The result of a calibration permits either the assignment of values of measurands to the indications or
the determination of corrections with respect to indications.
A calibration may also determine other metrological properties such as the effect of influence
quantities.
The result of a calibration may be recorded in a document, sometimes called a calibration certificate
or a calibration report.
[VIM: 1993 ISO International vocabulary of basic and general terms in metrology]
Certified Reference Material

Reference material, accompanied by a certificate, one or more of whose property values are certified by a
procedure, which establishes traceability to an accurate realisation of the unit in which the property values
are expressed, and for which each certified value is accompanied by an uncertainty at a stated level of
confidence.[ISO Guide 30:1992]
Limit of Determination
Applied to quantitative microbiological tests - The lowest number of micro-organisms within a defined
variability that may be determined under the experimental conditions of the method under evaluation.
Limit of Detection
Applied to qualitative microbiological tests- The lowest number of micro-organisms that can be detected,
but in numbers that cannot be estimated accurately.

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Negative Deviation
Occurs when the alternative method gives a negative result without confirmation when the reference
method gives a positive result. This deviation becomes a false negative result when the true result can be
proved as being positive.
Positive Deviation
Occurs when the alternative method gives a positive result without confirmation when the reference
method gives a negative result. This deviation becomes a false positive result when the true result can be
proved as being negative.
Reference Cultures
Reference strains, Collective term for reference strain, reference stocks and working cultures.
Microorganisms defined at least to the genus and species level, catalogued and described according to its
characteristics and preferably stating its origin.[ISO 11133-1:2000] Normally obtained from a recognized
national or international collection.
(Within India reference strain can be obtained from IMTECH, Chandigarh; National Chemical Laboratory,
Pune; Christian Medical College, Vellore; Central Research Institute, Kasouli, HP; National Institute of
Communicable Diseases, Delhi etc.)
Reference Material
Material or substance one or more of whose property values are sufficiently homogeneous and well
established to be used for the calibration of an apparatus, the assessment of a measurement method, or
for assigning values to materials.
[ISO Guide 30:1992]
Reference Method
Thoroughly investigated method, clearly and exactly describing the necessary conditions and procedures,
for the measurement of one or more property values that has been shown to have accuracy and precision
commensurate with its intended use and that can therefore be used to assess the accuracy of other
methods for the same measurement, particularly in permitting the characterization of a reference material.
Normally a national or international standard method.

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Reference Stocks
A set of separate identical cultures obtained by a single sub-culture from the reference strain. [ISO 111331:2000]
Relative Trueness
The degree of correspondence of the results of the method under evaluation to those obtained using a
recognized reference method.
Repeatability
Closeness of the agreement between the results of successive measurements of the same measurand
under the same conditions of measurement.
[VIM: 1993 ISO International vocabulary of basic and general terms in metrology]
Reproducibility
Closeness of the agreement between the results of measurements of the same measurand carried out
under changed conditions of measurement. [VIM: 1993 ISO International vocabulary of basic and general
terms in metrology]
Sensitivity (applied to microbiological tests)
The fraction of the total number of positive cultures or colonies correctly assigned in the presumptive
inspection. [ISO 13843:2000]
Specificity (applied to microbiological tests)
The fraction of the total number of negative cultures or colonies correctly assigned in the presumptive
inspection. [ISO 13843:2000]
Working Culture
A primary sub-culture from a reference stock. [ISO 11133-1:2000]
Validation
Confirmation, through the provision of objective evidence, that the requirements for a specific intended
use or application have been fulfilled. [ISO 9000: 2000]
Verification
Confirmation, through the provision of objective evidence, that specified requirements have been fulfilled.
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APPENDIX - C
METHOD VALIDATION
Laboratories with the appropriate knowledge, skills, experience and resources to do so in a competent
and thorough manner should only carry out validation of biological testing methods. The requirements for
method validation are detailed in Clause 5.4.5 of ISO/IEC 17025:2005.
The diagram on the following page (Figure 1) provides a very generalized approach to method validation
It is not intended to be a comprehensive reference to validation requirements, but rather a starting point to
assist laboratories to ensure the key components are considered. In some instances laboratories may
need to do more to demonstrate full validation; in other instances, some of the elements may not need to
be considered - depending on the purpose to which the method is to be applied.

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Customer requirements need to be defined and should Include but not be

limited to:
- Why is testing being done?
- Is there a specification limit?
- What accuracy is required?
- What detection limit/precision is required?
- Turn around time?
- Cost (including development)?
Define
Customer
Requirement
Validated
Method
Available?
No
No
No
Source a validated method from:

- International Standards
- National Standards
- Other Validated Methods e.g. ASTM, AOAC, AOCS, APHA etc.
Yes
Verify
Laboratory
Performance
Unvalidated
Method
Verify laboratory performance through:

- proficiency testing
- reference materials
- detection limit determination
- repeatability determination
- reproducability determination
- consumables verified
Unvalidated methods may be available from:
- journals
- customers
- in house
Validate
Method
Fit for
Purpose?
All methods need validation for example by:

- proficiency testing
- reference materials
- linearity confirmation
- specificity confirmation
- robustness assessment
- matrix effects/spiking
- detection limit / determination
- repeatability/reproducability determination
- consumables verified
Yes
If the method does not meet customer requirements then alternative methods
need to be sourced and verified/validated, and/or customer requirements
reviewed.
Document
Validation
Verification
Develop routine quality control programme: e.g.

duplicates
spikes
reference materials
proficiency testing
Following implementation a review programme should be instigated
Develop QC
Programme
Document
Laboratory
Method
Implement
Review

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APPENDIX - D
Reference Culture Maintenance, Subculture and Storage
Microorganisms have an inherent tendency to mutate in laboratory culture. It is essential then that
laboratories use procedures to maintain their cultures in a viable and genetically stable state. Various
methods have been established to preserve cultures so that minimum genetic drift occurs.
This section provides information to laboratories on the general principles involved on culture
maintenance. They are generally applicable to most aerobic organisms that are in common use. However
it should be noted that culture conditions for anaerobic organisms are significantly different and will
require suitable anaerobic environment. These organisms may be grown in anaerobic jars or chambers.
General rules for preparation of reference / working stock from reference culture
Reference culture is defined as a culture that is obtained from recognized microbial type culture collection.
Reference stock is the one that is derived from an authentic reference culture. A working stock culture is
the one that is derived from a reference stock culture and is used on a day-to-day basis in most of the
microbiological laboratories.
Reference cultures shall be sub-cultured only once to provide reference stocks. The reference stocks
must be used to prepare working stocks for routine works and working stock must not be refrozen and
reused once thawed. Working stocks shall not be sub cultured to replace reference stocks.
Microorganism Maintenance Plan
Tier-1
Reference culture received from culture collection
Tier-2
Reference stocks
Tier-3
Working stock (Daily QC use)

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Storage of reference organisms

Appropriate technique shall be used to preserve the reference microorganism so that the desired
characteristics of the strains are maintained. The laboratory shall assign suitable trained staff for
maintenance of reference culture. Reference culture can be stored by one of the following techniques:
Reference culture
Reference culture procured from culture collection can be stored in refrigerator (2C 8C) in the
original sealed vial or container till the expiry date. Reference culture once revived should be
stored as per the instructions provided by the agency or as per laboratory internal procedure.
Reference stock
The reference stock can be maintained at deep freezer (-18 0 C) or ultra cold freezer (-70C) for a
long storage (typically 2 years depending on individual culture viability). If the reference stock is
maintained at 4C on an appropriate medium, it can be used for a shorter time, typically up to
three months provided the culture viability is maintained. Some organisms used in antibiotic
testing lose their resistance over long storage hence advisable to prepare subcultures every 2
weeks. Many other tests require microorganisms not more than 24 hours old. Fastidious
organisms such as Streptococcus pneumoniae need to be subcultured every 3rd day. So
considering these specific cases, laboratory needs to decide their own storage plan which shall
be technically justifiable.
Working stock
Working stock cultures can be stored in refrigerator (2C 8C) and used on a day-to-day basis.
All aerobic bacteria can be stored in 2C 8C and used as per laboratorys subculture plan.
Anaerobic organisms should be stored in anaerobic conditions as per instructions provided by
culture collection or reference texts.
Guidance on preparation of frozen beads for long term storage

Freezing on bead is one of the culture preservation methods to prepare a reference stock culture
for long term storage. This method employs the drying of organisms from the liquid state on inert
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substrates, porcelain beads. These methods are suitable for short to medium term preservation at
-18C to - 70C for periods not exceeding one to five years respectively, with good genetic
stability.
The viability of the culture in frozen condition is species or strain specific. In general most of the
cultures being used for various testing purposes can retain good viability at -700c for 1-5 years
and at -180c for six months to one year. When it is observed that the culture is non viable , it may
be procured again from the reputed culture collections.
The procedure essentially consists of taking a pure culture from solid media and inoculating into a
suitably prepared vial containing appropriate broth medium and unglazed porcelain beads. After
agitating the beads in the broth, all excess fluid is removed from the vial with a fine tip Pasteur
pipettes. The vial is stored at -18C to -70C. Recovery is affected by removing a single bead
aseptically from the vial and inoculating it directly onto solid media or in to broth (Tier 3). The
remaining beads are available for later use.
Storage of reference cultures must be appropriately segregated from test samples.

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APPENDIX - E
Biosafety Levels
There are four levels of biosafety precautions for biological agents.
Biosafety level 1 is suitable for involving well characterized agents not known to consistently cause
disease in healthy adult humans and of minimal potential hazard to laboratory personnel and the
environment, work is generally practiced on open bench tops using standard microbiological practices.
Special containment equipment or facility design is neither required nor generally designed. Laboratory
personnel have specific training in the procedures conducted in the laboratory and are supervised by a
qualified and trained person in the area of microbiology or related science.
Biosafety Level 2 is similar to Biosafety Level 1 and is suitable for work involving agents of moderate
potential hazard to personnel and the environment. It differs from the level 1 by
1)
Laboratory personnel have specific training in handling pathogenic agents and are directed by
competent personnel.
2)
Access to the laboratory is limited when work is being conducted
3)
Extreme precautions are taken with contaminated sharp items and ;
4)
Certain procedures in which infectious aerosols or splashes may be created are conducted in
biological safety cabinets or other physical containment equipment
Biosafety Level 3 is applicable to clinical, diagnostic, teaching research or production facilities in which
work is done with indigenous or exotic agents which may cause serious or potentially lethal disease as a
result of exposure by the inhalation route. Laboratory personnel have specific training in handling
pathogenic and potentially lethal agents, and are supervised by competent scientists who are experienced
in working with these agents.
All procedures involving the manipulation of infectious materials are conducted within biological safety
cabinets or other physical containment devices or by personnel wearing appropriate, personal protective
clothing and equipment. The laboratory has special engineering and design features.
It is recognized however that some existing facilities may not have all the facility features recommended
for Biosafety Level 3 (i.e., double door access zone and sealed penetration). In this circumstance, an
acceptable level of safety for the conduct of routine procedures, (e.g., diagnostic procedures involving the
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propagation of an agent for identification, typing, susceptibility testing, etc.) may be achieved in a
Biosafety Level 2 facility providing:
1) The exhaust air from the laboratory room is discharged to the outdoors
2) The ventilation to the laboratory is balanced to provide directional airflow into the room,
3) Access to the laboratory is restricted when work is in progress and the
4) Recommended Standard Microbiological Practices, special practices and safety equipment for
Biosafety level 3 are rigorously followed.
The decision to implement Biosafety level 3 recommendations should be made only by the laboratory
director.
Biosafety Level 4 is required for work with dangerous and exotic agents that pose a high individual risk of
aerosol transmitted laboratory infections and life threatening disease. Agents with a close or identical
antigenic relationship to Biosafety Level 4 agents are handled at this level until sufficient data are obtain
either to confirm continued work or to work them at a lower level.
Members of the laboratory staff have specific and thorough training in handling extremely hazardous
infectious agents and they understand the primary and secondary containment functions of the standard
and special practices, the containment equipment and the laboratory design characteristics. They shall be
supervised by competent scientists who are trained and experienced in working with these agents. The
laboratory director should strictly control access to the laboratory.
The facility is either a separate building or in a controlled area within a building, which is completely
isolated from all other areas of the building. A specific operation manual is prepared or adopted.
Within work areas of the facility all activities are confined to Class III biological safety cabinets or Class II
biological safety cabinets used with one-piece positive pressure personnel suits ventilated by a life
support system.
The Biosafety Level 4 laboratory has special engineering and design features to prevent microorganisms
from being disseminated into the environment.

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APPENDIX- F
Uncertainty of Measurement
The approach is based upon overall variability of analytical process being conducted by the use of a
specific method in a particular laboratory. In addition to follow the test procedure without any deviation the
laboratory must have properly designed in house quality checks. The approach is required to meet the
underlying principles of the process.
(A) For each of the methods in the scope of accreditation providing numerical results the laboratory
should identify all components of the testing process which will contribute to the uncertainty in the final
results. At this stage it is not necessary to quantify each component but rather confirm its existence.
.Possible approaches for doing this exercise are:
(1) By critically evaluating each step in the documented method to identify those components that
may affect the results.
(2) By using the method equation and critically evaluating each variable to identify the components
that will affect its value.
(B) Identify and gather or collate all available data relating to the performance of the method. The source
of such data may be external to the laboratory or data generated internally.
(1) External data such as:
Published validation data for the standard method
Result from Proficiency testing or inter laboratory comparison programs.
(2) Internal data such as:
In house validation studies
Precision or repeatability data from duplicates
M/U values from calibration certificates
Variability in spike recovery data.
Standard/in house reference material results

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(C) Conduct a gap analysis to assess which of the component identified in (a) are incorporated in the
data collected in (b) It is important to have a clear understanding of how the data collected in (b) are
generated and what they mean. The following are a few examples which illustrate the type of issues that
need to be considered.
For numerical estimations a data from duplicate plating alone will not provide an adequate
estimation of measurement uncertainty as this is only a measure of individual ability to repeatable
plate and count. It will not include the majority of other major components of uncertainty such as
sample preparation, initial dilutions, diluents, dilution equipment, media performance and
incubation etc.
In case data is being taken from duplicate sample testing without taking note of other variables
being identified in the first step, will only include the component associated with taking the test
portion from the submitted test sample and will not normally include the component of uncertainty
associated with different equipments, different operators, different batches of media and reagents
etc. The precision data from duplicates would in itself give an under-estimation of the overall
uncertainty.
In case if as many of the testing variables that are the components identified in (a) are being
incorporated in the analysis of each duplicate of each sample such as different analysts, different
batches of diluents, media reagents, different pipettes, incubators, ovens etc, then this data
(another form of intermediate precision) will provide a more realistic assessment of measurement
uncertainty. For many biological testing laboratories where the received sample is not stable, this
approach may be the only realistic one to estimate measurement uncertainty.
The precision data from true duplicate sample testing collected over a long period of time in which
the identified components were varied, may provide a possible estimate of uncertainty.
Appropriate statistical analysis may be based upon the data generated in intra or inter laboratory
collaborative studies on the use of a method based upon intermediate precision to analyze a
diversity of matrices.
Intermediate precision data from reference materials analyzed repeatedly over time would include
the components associated with different analyst, media equipment and reagents etc .However

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by their nature the reference materials are homogeneous and stable and thus this intermediate
precision data will not include the components associated with sub sampling the test portions
from real test samples.
Reproducibility data would generally give an over estimation of an individual laboratory

uncertainty of measurement as it include many different analysts, type of equipments, batches of
media and often different methods and some of these components are not relevant to a particular
laboratory circumstances. In most of the cases such
data is normally
generated from
homogeneous and stable samples and that may not reflect actual practices in working
laboratories.
Spike recovery data needs to be carefully considered. The actual recovery itself is not a
component of measurement uncertainty as it can be corrected for. Spiking may also be required
in case intermediate precision approach is to be considered in order to obtain statistical significant
counts.
(D) Where there are components identified in (a) are not incorporated in data collected in (b) these needs
to be independently estimated and their significance assessed.
If they are significant the laboratories will need to review and re design their quality control data collection
programs in order to incorporate as many of these additional components of uncertainty as possible.
Components of uncertainty which cannot be incorporated in to the quality control data generated can be
estimated by separate experiment, from published data, from calibration certificates, certificates of
analysis or by professional judgment.
Depending upon the principles addressed in the above mentioned approach the laboratories should be
able to obtain data to sufficiently cover all significant identified components of uncertainty coming from
different sources.
The approaches suggested above will generally provide appropriate consideration of all these issues and
result in a reasonable estimate provided the data are generated from same or similar matrix.

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Points of Consideration
In the vast majority of the tests of biological nature the result is dependent on the method being
used. If the method is followed the method bias does not contribute to the measurement
uncertainty. The best estimate of the uncertainty of measured results will therefore come from the
uncertainty associated with the performance of the method used.
In quantitative biological testing it is ideal if the uncertainty estimation is evaluated at selected

levels across the range application of the method. Often a test is conducted to assess compliance
with a particular specification /regulatory limit etc. In these instances laboratories should at least
estimate uncertainty value attributable to measurement results close to the specification limits.
For quantitative determinations the laboratories are reminded that results from plate count tests
have a skewed distribution and thus require log transformation to approximate normal distribution
statistics. Log standard deviation /confidence limits should then be calculated before anti logging
each limit independently
For tests involving MPN methods where test results are obtained from relevant tables, the
significant component of uncertainty are already built in to MPN table values. For MPN results the
values from the 95% confidence columns of the tables are being accepted as a reasonable
estimates of uncertainty of these results provided that laboratory follows the test methods and
subsequent reporting instructions along with the assurance that laboratory estimates of precision
(duplicate assays) fall within these values.
It is recognized that in some instances the approaches addressed in the procedure may take
some time for its implementation. The laboratory may require to re design their in house quality
checks program because the data may need to be collected over a reasonable length of time in
order to make a sufficiently rigorous assessment of measurement uncertainty. The laboratories
are required to maintain records of each test or type of tests to demonstrate full implementation of
the required procedure

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Uncertainty of measurement is one of the most difficult parameters to establish in GMO detection.
Variance associated with the sampling step is likely to constitute the major contribution to the
overall variance, since GMOs are usually non-homogeneously distributed in the bulk. The
biological diversity of a particular GMO, such as different zygosity or ploidy can be another source
of variance. The laboratory has no control over such variables but must be aware of them for the
correct interpretation of results. Quantitative PCR methods measure DNA%, if this needs to be
converted into copy numbers, weight%, GMO%, the factors listed above and the one listed in Lipp
et al. 2003 will apply and need to be factored into the MU.
In the testing laboratory, additional MU can result from in adequate sample homogenization,
resulting in differences in GMO content between test portions taken for DNA isolation. The
performance of the system of homogenization used is checked in due course by independent
analysis of test portions.
Additional uncertainty is added in subsequent PCR analysis due to MU of the endogenous

reference and of transgene copy number determination. The standard deviation of parallel PCR
quantity estimations increases when the target copy numbers
in PCR reaction are low, resulting in high final MU for samples with low target DNA content.
Parallel real-time PCR measurements must be performed to allow statistical evaluation of the
variability introduced by the PCR analysis.

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REFERENCES
1. ISO/IEC 17025:2005, General Requirements for the Competence of Testing and Calibration
Laboratories.
2. ISO 7218:2000, Microbiology of Food and Animal Feeding Stuffs - General Rules for
Microbiological Examinations.
3. ISO 6887-1:1999 Microbiology of Food and Animal Feeding Stuffs - Preparation of Test
Samples, Initial Suspension and Decimal Dilutions for Microbiological Examination. Part 1 General Rules for the Preparation of the Initial Suspension and Dilution.
4. ISO Guide 30:1992, Terms and Definitions Used in Connection with Reference Materials.
5. ISO 9000, Quality Management Systems - Fundamentals and Vocabulary.
6. VIM: 1993, International Vocabulary of Basic and General Terms in Metrology.
7. ISO (CIPM):1995, Guide to the Expression of Uncertainty in Measurements
8. Draft ISO/DIS 16140, Microbiology of Food and Animal Feeding Stuffs- Protocol for the
Validation of Alternative Methods.
9. ISO 13843:2000, Water Quality Guidance on Validation of Microbiological Methods.
10. ISO 11133-1:2000, Microbiology of Food and Animal Feeding Stuffs. Guidelines on
Preparation and Production of Culture Media. Part 1- General Guidelines on Quality
Assurance for the Preparation of Media in the Laboratory.
11. Draft ISO/FDIS 11133-2, Microbiology of Food and Animal Feeding Stuffs. Guidelines on
Preparation and Production of Culture Media. Part 2- Practical Guidelines on Performance
Testing on Culture Media.
12. EN 12741, Biotechnology- Laboratories for Research, Development and Analysis Guidance
for Biotechnology Laboratory Operations.
13. ISTA Seed Testing Laboratory Accreditation Standard Version 3.1
14. EA-04/10:2002 Accreditation for Microbiological Laboratories
15. HOKLAS Supplementary Criteria No.8
16. HOKLAS Supplementary Criteria No.21
17. IANZ Specific Criteria for Accreditation -Biological Testing
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18. Guide for and
the care use of
Laboratory Animals, Institute of Laboratory Animal
Resources, Commission on Life sciences, National Research Council,Washington,DC,1996

19. INSA Guidelines for care and use of Animals in Scientific Research, New Delhi, 1992
20. CPCSEA guidelines for Laboratory Animal Facility
21. Laboratory Animal Management, Series on Rodents and Dogs, National Research Council,
Washington, DC, 1996.
22. Guide to the expression of Uncertainty in Measurement, First edition ISO-1995.
23. Eurachem guide: Quantifying Uncertainty in analytical measurements 2nd edition 2000.
24. APLAC TC 005- Interpretation and guidance on the estimation of uncertainty of
measurements in testing.
25. Nordic committee on food analysis (NMKL) Procedure No-08, Measurement of uncertainty in
microbiological examination of food (1999).
26. ASTG5, Uncertainty of measurement, precision and limit of detection in chemistry and
microbiology.
27. Uncertainty of quantitative determinations derived by cultivation of microorganisms, published
by advisory commission for metrology, Finland, 2002.
28. Microbiology of food and animal feeding stuff-Guidelines for the estimation of measurement
uncertainty for quantitative determinations, ISO/TS-19036: 2006(E).
29. Miraglia; K.G. Berdal, C. Brera, P. Corbisier, A. Holst-Jensen, E.J. Kok, H.J.P. Marvin, H.
Schimmel, J. Rentsch, J.P.P.F. van Rie, J. Zagon (2004) Detection and traceability of
genetically modified organisms in the food production chain. Food Chem Toxicol 42:1157
1180.
30. Markus Lipp (2003) Testing for foods derived from modern biotechnology: opportunities and
limitations for metrology. Accreditation and Quality Assurance: Journal for Quality,
Comparability and Reliability in Chemical Measurement 8:454460.
31. Trapmann et.al, (2007) European Commission, Directorate General-Joint Research Centre,
Guidance document on Measurement Uncertainty in GMO testing Laboratories- EUR report
22756 EN, 1-46.
32. ISO/IEC 17043:2010 Conformity Assessment General requirements for Proficiency Testing

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COMPOSITION OF THE TECHNICAL COMMITTEE

Shri S.K. Gaind (Chairman)
Technical Advisor
Avon Food Lab Pvt Ltd
Delhi 110 035
Dr. Deepak K. Agarwal
Former Scientist-G
Industrial Toxicology Research Centre
P.O. Box 80, Mahatma Gandhi Marg
Lucknow
Dr. R. Pirabhakaran
Principal Scientist-Microbiology
CavinKare Research Centre
Chennai
Dr. P.K. Dhakephalkar
Scientist, Microbial Sciences Division
Agharkar Research Institute (ARI-DST)
Agharkar road, Pune
Dr. Gurinder Jit Randhawa
Principal Scientist
National Research Center for DNA fingerprinting
National Bureau of Plant Genetic Resources (NBPGR)
Pusa, New Delhi
Dr. .G. S. Prasad
Scientist-F, Microbial Type Culture Collection
Institute of Microbial Technology (IMTECH)
CSIR, Chandigarh
Dr. Anand Vally Amma
WHO/FAO Consultants
Cochin
Shri Vijender P. Gupta
Head, Nestle Quality Assurance
Centre for South Asia Region
Nestle India Limited, Moga
Shri. Anil Relia
Director, NABL
Mrs. Anuja Anand,
Accreditation Officer-II, NABL
Mr. Anand Deep Gupta,
Accreditation Officer-I, NABL

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3rd Floor, NISCAIR
14, Satsang Vihar Marg
New Mehrauli Road
New Delhi 110 067
Tel.: +91-11 46499999
Fax: +91-11 26529716
Website: www.nabl-india.org

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NABL 102

National Accreditation Board for Testing and Calibration Laboratories

Specific Criteria for Biological Testing Laboratories

Introduction and Scope of Document

Accommodation and Environment

Test Methods and Method Validation

Equipment - Maintenance, Calibration and Performance

Disposal of Contaminated Waste

Assuring Quality of Test Results

Reporting the Results

Appendix A Classes of Tests in Biological Discipline

Appendix B Glossary of Terms

Appendix C Method Validation

Appendix D Reference Culture Maintenance, Subculture and Storage

Appendix E Bio-safety Levels

Appendix F Uncertainty of Measurement

Composition of the Technical Committee

National Accreditation Board for Testing and Calibration Laboratories

Specific Criteria for Biological Testing Laboratories

Introduction and Scope of Document

As majority of accredited biological testing laboratories are primarily involved in bacteriological

in biological testing pertaining to

National Accreditation Board for Testing and Calibration Laboratories

Specific Criteria for Biological Testing Laboratories

Personnel (ISO/IEC 17025 clause 5.2)

Authorized signatory should fulfill either of the following requirements :

National Accreditation Board for Testing and Calibration Laboratories

Specific Criteria for Biological Testing Laboratories

National Accreditation Board for Testing and Calibration Laboratories

Specific Criteria for Biological Testing Laboratories

Accommodation and Environment (ISO/IEC 17025 Clause 5.3)

Sample receipt and sample storage

Media preparation and Sterilization

Storage of reference cultures/Reference materials

Decontamination and washing

National Accreditation Board for Testing and Calibration Laboratories

Specific Criteria for Biological Testing Laboratories

Concentrated infectious agents such as enriched cultures of pathogenic

National Accreditation Board for Testing and Calibration Laboratories

Specific Criteria for Biological Testing Laboratories

The laboratory shall have pest control programme /schedule.

Additional Requirements for GMO Testing Laboratories

Reagents, consumables and equipment shall be located at appropriate designated areas

National Accreditation Board for Testing and Calibration Laboratories

Specific Criteria for Biological Testing Laboratories

General Requirements for Toxicology Laboratories

National Accreditation Board for Testing and Calibration Laboratories

Specific Criteria for Biological Testing Laboratories

Environmental temperature and humidity conditions of animal house should be

Adequate ventilation, necessary for adequate supply of oxygen, is generally considered

National Accreditation Board for Testing and Calibration Laboratories

Specific Criteria for Biological Testing Laboratories

cycle is generally acceptable. A time-controlled lighting system should be used to ensure

Specific Criteria for Biological Testing Laboratories

National Accreditation Board for Testing and Calibration Laboratories

Specific Criteria for Biological Testing Laboratories

Other test systems including tissues, cells, sub-cellular preparations, micro-organisms,

vials/tubes/receptacles/containers. Since the labels on such small containers may not

National Accreditation Board for Testing and Calibration Laboratories

Specific Criteria for Biological Testing Laboratories

Test Methods and Method Validation (ISO/IEC 17025 clause 5.4)

Lab developed/non standard methods