Preparation For Diagnosing Infections

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Immunology Applications > Preparation for Diagnosing Infections
Preparation for Diagnosing Infections
Specimen Collection
Immediate Direct Examination of Specimen
Cultivation of Specimen
DNA Analysis Using Genetic Probes and PCR
Nucleic Acid Sequencing and rRNA Analysis
www.boundless.com/microbiology
Specimen Collection
Specimen collection requires withdrawing blood, cerebrospinal fluid, collecting
urine, or swabs from mucosal surfaces.
Specimen collection is performed using aseptic techniques to ensure sterility of
the sample and avoid contamination from bacteria or other bodily fluids.
The types of biological samples accepted in most clinical laboratories are: serum
samples, virology swab samples, biopsy and necropsy tissue, cerebrospinal fluid,
whole blood for PCR, and urine samples.These are collected in specific
containers for successful processing in the laboratory.
Venipuncture
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www.boundless.com/microbiology/textbooks/boundless-microbiology-textbook/immunology-applications-12/preparation-for-diagnosing-infections-1
49/specimen-collection-753-5428
Immediate Direct Examination of Specimen

Immediate direct examination methods depend on the nature of the specimen.
Diagnostic laboratory techniques include direct testing using a microscope, and
immunological or genetic methods that provide immediate clues as to the identity
of the microbe or microbes in the sample.
Phenotypic methods include the examination of microscopic, macroscopic, and
biochemical characteristics of a pathogen.
Microscopic examination of E. coli

49/immediate-direct-examination-of-specimen-754-4901
Cultivation of Specimen
The success of pathogen identification and treatment depends on how the
specimen is collected, handled, and stored.
Diagnostic laboratory techniques include direct testing using a microscope, and
immunological or genetic methods that provide immediate clues as to the identity
of the microbe or microbes in the sample.
Following direct testing, cultivation, isolation, and identification of pathogens
using a wide variety of general and specific tests is required.
Blood culture
9/cultivation-of-specimen-755-722
DNA Analysis Using Genetic Probes and PCR

Hybridization can identify a bacterial species by analyzing segments of its DNA.
Genetic probes are small fragments of DNA or RNA that are complementary to
the specific sequences of DNA from a particular microbe.
This approach is most useful in the detection of infections due to microorganisms
that are difficult to culture.
Mycobacterium Tuberculosis
49/dna-analysis-using-genetic-probes-and-pcr-756-5430
Nucleic Acid Sequencing and rRNA Analysis

This method is effective in identifying general group differences of pathogens.
16S rRNA gene sequences contain hypervariable regions that can provide
species-specific signature sequences useful for bacterial identification.
This method can also be fine-tuned to identify pathogens at the species level.
Ribosomal RNA
9/nucleic-acid-sequencing-and-rrna-analysis-757-8429
Appendix
Immunology Applications
Key terms
biopsied to remove and examine a sample of tissue from a living body for diagnostic purposes.
biopsy The removal and examination of a sample of tissue from a living body for diagnostic purposes.
MacConkey agar A culture medium designed to grow Gram-negative bacteria and differentiate them from lactose fermentation.
mannitol A polyhydroxy alcohol, an isomer of sorbitol, used as an artificial sweetener.
necropsy The pathological dissection of a corpse; particularly to determine cause of death.Applicable to the examination of
any life form.
PCR polymerase chain reaction
polymerase chain reaction A technique in molecular biology for creating multiple copies of DNA from a sample; used in genetic
fingerprinting etc.
ribosomes Large and complex molecular machine, found within all living cells, that serves as the primary site of biological
protein synthesis.
Blood culture
This blood is cultured in a bottle to detect bloodstream infections.
Wikipedia. "Blutkultur - blood culture." CC BY-SA http://en.wikipedia.org/wiki/File:Blutkultur_-_blood_culture.jpg View on Boundless.com
Microscopic examination of E. coli

Gram negative E. coli examined under the microscope from a patient with urinary tract infection.
Wikipedia. "E choli Gram." CC BY-SA http://en.wikipedia.org/wiki/File:E_choli_Gram.JPG View on Boundless.com
Mycobacterium Tuberculosis
a Mycobacterium tuberculosis culture revealing the organism's colonial morphology.
Wikipedia. "TB Culture." CC BY-SA http://en.wikipedia.org/wiki/File:TB_Culture.jpg View on Boundless.com
Ribosomal RNA
Structure and shape of the E.coli 70S ribosome.The large 50S ribosomal subunit (red) and small 30S ribosomal subunit (blue) are shown with a 200
ngstrom (20 nm) scale bar. For the 50S subunit, the 23S (dark red) and 5S (orange red) rRNAs and the ribosomal proteins (pink) are shown.For the
30S subunit, the 16S rRNA (dark blue) and the ribosomal proteins (light blue) are shown.
Wikipedia. "Ribosome shape." CC BY-SA http://en.wikipedia.org/wiki/File:Ribosome_shape.png View on Boundless.com
What is the preferred specimen source for serologic testing?
A) plasma
B) serum
C) cerebrospinal fluid
D) platelets
What is the preferred specimen source for serologic testing?
A) plasma
B) serum
C) cerebrospinal fluid
D) platelets
Boundless - LO. "Boundless." CC BY-SA 3.0 http://www.boundless.com/
Which of the following IS NOT a useful trait when determining

specimen identification?
A) Gram-stain reaction
B) acid fast reaction
C) special structures
D) nutritional activity
Which of the following IS NOT a useful trait when determining

specimen identification?
A) Gram-stain reaction
B) acid fast reaction
C) special structures
D) nutritional activity
Direct fluorescence antibody tests can:
A) diagnose infection
B) indicate the presence of antibody
C) indicate the presence of complement
D) determine the oxygen requirements of a microorganism
Direct fluorescence antibody tests can:
A) diagnose infection
B) indicate the presence of antibody
C) indicate the presence of complement
D) determine the oxygen requirements of a microorganism
In order to identify microorganisms that are difficult to culture, the

microorganisms' DNA must first be:
A) labeled
B) hybridized to an array of immobilized diagnostic probes
C) digested with restriction enzymes
D) amplified by PCR
In order to identify microorganisms that are difficult to culture, the

microorganisms' DNA must first be:
A) labeled
B) hybridized to an array of immobilized diagnostic probes
C) digested with restriction enzymes
D) amplified by PCR
Which of the following has been most useful in uncovering

evolutionary relationships between different organisms?
A) cytochrome c
B) 23S rRNA
C) 5S rRNA
D) 16S rRNA
Which of the following has been most useful in uncovering

evolutionary relationships between different organisms?
A) cytochrome c
B) 23S rRNA
C) 5S rRNA
D) 16S rRNA
Attribution
Wikipedia. "Blood culture." CC BY-SA 3.0 http://en.wikipedia.org/wiki/Blood_culture
Wikipedia. "MacConkey agar." CC BY-SA 3.0 http://en.wikipedia.org/wiki/MacConkey%20agar
Wiktionary. "mannitol." CC BY-SA 3.0 http://en.wiktionary.org/wiki/mannitol
Wikipedia. "Blood samples." CC BY-SA 3.0 http://en.wikipedia.org/wiki/Blood_samples
Wiktionary. "biopsied." CC BY-SA 3.0 http://en.wiktionary.org/wiki/biopsied
Wikipedia. "Diagnostic immunology." CC BY-SA 3.0 http://en.wikipedia.org/wiki/Diagnostic_immunology
Wiktionary. "PCR." CC BY-SA 3.0 http://en.wiktionary.org/wiki/PCR
Wiktionary. "biopsy." CC BY-SA 3.0 http://en.wiktionary.org/wiki/biopsy
Wiktionary. "necropsy." CC BY-SA 3.0 http://en.wiktionary.org/wiki/necropsy
Wikipedia. "Polymerase chain reaction." CC BY-SA 3.0 http://en.wikipedia.org/wiki/Polymerase_chain_reaction
Wikipedia. "DNA profiling." CC BY-SA 3.0 http://en.wikipedia.org/wiki/DNA_profiling
Wiktionary. "polymerase chain reaction." CC BY-SA 3.0 http://en.wiktionary.org/wiki/polymerase+chain+reaction
Wikipedia. "16S ribosomal RNA." CC BY-SA 3.0 http://en.wikipedia.org/wiki/16S_ribosomal_RNA
Wikipedia. "ribosomes." CC BY-SA 3.0 http://en.wikipedia.org/wiki/ribosomes

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Immunology Applications > Preparation for Diagnosing Infections