Microbiology technique

|Phagogenisis | Mycology
| Urine Analysis | Gram
Positive Bacteria |
Enteric Bacteria |
Antibiotics | PCR |
Serology |Fussy bacteria|
Parasitology | Virology |
Epidemology
Phathogenesis

Virulence Factor:
Factors that involve,Adherence
,Invasion,Evasion, Toxigenicity

Adherence Adhesins
Adhesion/Attachment of bacteria
cell is the first process of Exopolysaccharide
invasion .It is nessacery to
prevent bacteria cells from being Lipoteichoic acid
swept away.
Invasion Flagella
The process by which a
pathogen , enters and spread in
tissue
Evasion Exopolysaccharide capsule
Allows the bacteria to avoid Immunoglobin Proteases
damage from host cell immune Complement Proteases
mechanisms . Eg Flagella
Antigen Switching
Neiserria Gonorrhea ,

Direct toxigenicity Exotoxins

Acts by interfering or harming
Cellular physiology though,
inhibition or destruction. Endotoxins

Indirect toxigenicity

Infection site Specimen collected

respiratory tract Sputum
Systemic infection blood specimen
brain and spine Cerebrospinal fluid
GI tract Stool
Urinary tract Urine
Genital tract Genital swabs
Skin and Mouth swab
Neiserria gonorrhea Streptococcus pneumoniae Clostridium difficile

Clostridium perfringens Staphylococcus aureus E. coli

Staphylococcus saprophyticus Proteus mirabilis Candida albicans

Yeast

Klebsiella species Staphylococcus aureus E. coli

Mycology
Mould(Multicellular Fillamentous Yeast
Fungi) Single cell
Cell wall made of Chitin Chitin cell wall
Formed from branching Hyphae Can have Pseudohyphae which linkes colonies
Cottony colonies Opaque cream-colored colonies,budding

Localization
Location Systematic Subcutaneous and Superficial
 Subcutateous
Description Infects the internal Found on nails, and deep Found on body surfaces,
organ , spread by layers of skin. vaginal surface, and
Dermis hair(?),dead layer of skin.
Spread by localization ,epidermis,hair,nails, Spread by skin contact
of opportunistic subcutis
pathogen and Infects after skin
proliferative growth. penetration or cut
Inhallartion
Etiology Histoplasma and Mycetoma, Sporotrichosis Tinea Pedis, Vaginal
Candida Candidiasis
Sample Blood for blood culture NailClipping Skin infection-Skin scrappings
Collection (more suitable for yeast)
or antigen detection
(cryptococcal and
Histoplasma antigen)
Pneumonia-Sputum,
bronchial biopsy?
Pathogenic Factors
Dimorphism
Glycoprotein Adherence
Capsules Resist Phagocyrtosis
Enzymes Keratinase, Elastase and Collagenase, Exotoxin Commented [DK1]: Toxin???
Proteinase
Toxins Alfatoxin

Microscopy (Morphology , Quick , Direct Evidence)

Wet Mount PBS Lactophenol Gram Stain
Cottonblue

Useful to detect Candida,

Cryptococcus and Aspergillus

Potassium Hydroxide India Ink Calcofluor white

Clearing of specimen to make For detection of C. Stains the cell wall of fungi,
fungi more readily visible; neoformans in CSF causing the fungi to fluoresce.
allows rapid detection of fungal Calcofluor white can be mixed
elements with potassium hydroxide for
the clearing of specimen to
make fungi more readily
visible.

Giemsa Germ-tube test

For detection of Histoplasma test for C. albicans. Germ Tube

capsulatum and
Pneumocystitis jirovecii
solutions contains tryptic soy
broth and fetal bovine serum. Pos
Test: Extension of germ tube. Neg
Test No Extension
Candida albicans produces germ
tube when incubated in serum at
37C for 2 to 4 hours.

Culture Media(Mycology)
Sabouraud dextrose agar Blood agars
 contains glucose and beef extract (pH 5.0) as Used to promote growth of yeast phase for
it does not support growth of bacteria. dimorphic fungi
Antibacterial antibiotics (gentamicin,
chloramphenicol and cycloheximide) added for
recovery of medical fungi from nonsterile
specimens

Inhibitory Mold Agar Sugar fermentation

contains chloramphenicol, is a moderately Soome fungi can only metabolize specifi sugars ,
selective medium used for the isolation of thus we can taRGET AND DETECT THE SPECIFIC
pathogenic fungi, exclusive of dermatophytes. DRUGS. yay

Serological
Molecular
Biochemical
Histopathological Haematoxylin and Eosin stain
Periodic acid Schiff stain
2. Direct Immunoflurescence
Classification
Zygomycota Ascomyota Basidiomycota Chytridiomycota
Produce Musroom
unspeted
Fungi
Reproduce by
sporangia
Mycelium
extend the area in which a fungi can find
nutrients
Large group of hyphae
Hyphae
The cells of hyphae have thin cell walls made
up of chitin. allow materials to flow among the
cells, and it enables the sharing of water and
nutrients absorbed from one part of hyphae
with other regions of the fungal
Sporangiospore
Sporangiophores are specialized aerial
hyphae that bear specialized sacs called
sporangia /sporangium
Sporangium
Produce spores
Conidiophore
Rhizoid
Anchor the Fungus
Stolon,
Connect hyphae together, from which rhizoid
and Sporangiospore and hyphae arise
 Urine Analysis
Pathogen Staphylococcus Proteus Candida Klebsiella
saprophyticus mirabilis albicans species
Gram(+) Yeast

Gram(+) Gram (-) Gram(-)

Staphylococcus Oxidase(-) Oxidase(-)
Facutative Facutative Facutative
Anaerobes Anaerobes Anaerobes
Enterobacteriacidae Enterobacteriacidae
coccus-shaped
Lactoase(-)
Bacilli shaped
Commenly cause
UTO Deaminate
Has Adhesins for Phylenalanina and
the epithelium are motile

Inhibited by CLED
medium and
Phenyethyl alcohol,
found on kidney
stones and hydrolyse
urea into ammonia
making urine alkaline
Etiology May originate from the bowl and occationally lymphatic or blood flora.
Sample Collection
Pediatric Collection bag attached to uretha, and urine is collected
Collection
method
Subprapubic Inserted to bladder though abdominal wall
Cathether Insertion of a carther into bladder
Midstream Recommended for microbiological culture and antibiotic subceptibility.
Midstream is collected as the midtream is less likely to have contaminants from
the uretha but the bladder or UT.
Transportation Kept in 4degC for 24 hours
Microscpic test
Centrifuge 12 mL of well-mixed urine in a conical centrifuge tube and dispose supernatant, leaving 1 mL load
sedinments in counting chamber . View under phase contrast with various magnifications Phase contrast used as
it give more details to the 3D casts.
Casts Waxy Cast Advanced Kidney diesease

Hyaline Dehydration and Diuretics

WBC Caused by Pyelonophitis and Glomerulephritis

Squamous Contamination of the Urine sample or damage

Epithelial to tubles in the kidney.
Cast Commented [DK2]: Minimum amount to indicate disease

composed of renal epithelial cells.seen in Commented [DK3R2]:

Renal
Tubular conditions such as renal tubular necrosis,
viral disease (such as CMV nephritis), and
Epithelial
kidney transplant rejection.
Cell Casts

Granular result either from the degeneration of

Casts cellular casts, or direct aggregation of
plasma proteins or immunoglobulin light
chains.They are seen after sternous
exercise, chronic renal diseases, acute
tubular necrosis etc
Microscpic test
Centrifuge 12 mL of well-mixed urine in a conical centrifuge tube and dispose supernatant, leaving 1 mL load
sedinments in counting chamber . View under phase contrast with various magnifications Phase contrast used as
it give more details to the 3D casts.
Crystal Calcium occur in urine of any pH. The crystals vary in size
Oxalate from quite large to very small. Dietary asparagus
Dihydrate and ethylene glycol intoxication are notorious for
Square urinary calcium oxalate formation.

Triple Associated with Proteous , Klebsilla , Seen

Phosphate in UTI patients with infections caused by
(Coffin urea splitting bacteria.
rectangular) Also called magnesium ammonium phosphate
crystals and form in abundance of ammonia

Uric Crystal Caused by reduced Kidney function,or no clinical value>

Cystine Cystinuria
Crystals
Amphorius No significnce
Bacteria
WBC High number sindicate pyuria and bacteria infextion due to
inflammation
RBC Increased RBC in urine is termed
hematuria, which can be due to
hemorrhage, inflammation, necrosis,
trauma or neoplasia somewhere
along the urinary tract (or urogenital
tract in voided specimens).
Contamination is commonly seen
when the patient is menstruating. Thus, collection of samples during
this period should be avoided.

Macroscopic
Culture Media
Blood Agar Mc Conkey

Differential Enriched Differential Selective

Dertermine Haemolytic To Cultivate fastidious Acid fermenters Gram (-)
properties bacteria Bile Salts : Inhibit Proteus and Gram(+)
Crystal Violet inhibits Gram(+)
5-10 percent Sheep/horse blood. Neutral Red Dye :
Beta Alpha Gamma Pink Red Yellow:
Total clear Greenish halo Lactose Neutral Basic non
zone due to due to No clearing or fermenting lactose
complete biliberdin , no/very little E. coli*** fermenting.
haemolysis partial haemolysis. Enetrobacter* Salmonella
haemolysis Klebsiella* Proteus
Streptococcus haemolyticus Citobacter** Yersinia
Pseudomonas
Aeruginosa
Shigella
*Klebsiella and Enterobacter
leads to mucoid colonies
**Citobacter is slow fermenting
*** Enterohemoregic E.coli is
dffrentiated using sorbitol.
Used to isolate Enteric bacteria

CLED Agar Colistin-Nalidixic Acid Agar

Electrolyte(-) : Prevents proteus swarming Collistin breaks cell membrane of Gram(-)
Cystine (+) : Promote cys Nailidixic block dna replication
Bromothymol Blue : Differential Selective
Acid / Basic/ Acid fermenters Inhibits (Gram-)
Lactoacillus L Cystine Enterobacteriacidae
Decarboxylation and pseudomonas
Allows the growth of
Yeast, Stapphylococci,
E.Coli : Opaque and yellow streptococci and
Klebsiella .spp , Yellow/whit mucoid in picture pneumococci.
Proteus: transparent Blue Corn starch promotes Neisseia.Spp
Pseudomonas : Green with matted surface NaCl supplies electrolytes for transport and
Enterococci : Small Yelllow Colonies osmotic balance
Staphylococcis : uniform Deep Yellow colonies Blood to determine haemolysis
Coagulase(-) Staphylococci , Pale Y ellow Beta Alpha Gamma
 Pathology
Terms Meaning Signs
Cystitis Infection of the urinary bladder - Especially common in women ->
- short distance of urethra to anus
-> pathogenic bacteria able to
travel up to bladder via adhesive
molecule

Bacteriuria bacteria in the urine

Asymptomatic Asymptomatic bacteriuria in the absence
bacteriuria of symptoms of pyuria or presence of
white blood cells on microscopic
examination.
Pyuria White blood cells in urine
Complicated UTI UTI occurring in functionally or - Occurs when GI bacteria spread
structurally abnormal urinary tract a host from the anus to the urethra
with a compromised immune system, a - Urethra close to vagina ->
host with diabetes, or infection with a herpes/ gonorrhea can cause
virulent or resistant strain of bacteria urethritis
-
Asymptomatic Bacteriuria without symptoms.
bacteriuria
Uncomplicated UTI occurring in a normal urinary tract in
UTI a normal person
Pyelonephritis Infection of the kidney

Urine Dipstick (First morning midstream?)

pH Determine the acidity of the Urine.

Proteus Mirabus produces urease , which makes urea turn

into ammonia, making the urine alkaline.

Leu -Test for WBC either whole or lysed

-If (+) = infection or inflammation of Urinary tract
-If (-) = infection is unlikely
NIT A positive nitrite test indicates that bacteria may be present in
significant numbers in urine. It is a
surrogate marker of bacteruria (presence of bacteria in urine)
E.g. Gram negative rods such as E. coli are more likely to
give a positive test.
PRO It tells us the protein excretion gm/24 hours Negative: <0.1
A color matching any block greater than 'Trace' indicates
mg/24hours
significant proteinuria (excess protein).
Elevated results indicate abnormal increased permeability of Tr(trace): 0.1-0.2 mg/24
the glomerulus membrane. Causes may be due to hours
pregnancy, 1+: 0.2-0.5 mg/24 hours
chemical poisons and more.
2+: to 0.5-1.5 gm/24 hours
3+: to 2-5 gm/24 hours
GLU Purpose : detect presence of glucose in the urine. (As we are
not supposed to urinate out glucose.)
Normal range : 0-0.8
If it exceeds the normal range, it could indicate health issues
such as diabetes
KET Purpose : detect presence of ketone bodies.
Small: <20 mg/dL
Moderate: 30 to 40 mg/dL
Large: >80 mg/dL
URG Intestinal bacteria convert bilirubin to
Urobilinogen(absorbed by kidneys)
Normal level less than or equal to
1 mg/dL
BIL Comes from the breakdown of
Haemoglobin in RBCs.
Even minute amounts of bilirubin indicates jaundice, liver
cirrhosis , hepatitis.
ERY Able to detect 5 to 250 RBC
- Normal range of ERY: > 3 ery/uL
Disorder : Hematuria (Blood in your urine)
- Causes: Kidney, urinary tract damage, bladder calculi
HB purpose: Test for the presence of haemoglobin in urine
- Able to detect 10 -250 RBC which lyse
Disorder: Hemoglobinuria( Haemoglobin in urine)
- Burns, Malaria, Hemolytic uremic syndrome
Gram Positive Bacteria
Staphylo aureus Clostridium Perfingens
Sperical cells
Found commensally on the skin and mucosal
surfaces especuially S Epidemidides

Some pathogenic staphylococcus aureus cause

blood haemolysis and blood coagulation.

Catalase positive , Coagulase (+)

Listeria Monocytogenes Bacillus Cerus

Enteric Gram Negative Bacteria {Facultive anaerobes or aerobes} Fermentative catalase (+) Oxiase
Negative(-)
Campylobacter Vibrio Spp
Enterobacter - Salmonella Enterobacter E. coli
Oathogenic and acquired though the oral route Indole (+)
H2S(+) Mannitol(+)
Lysine Decarboxylase(+)
Enterobacter - Shigella Klebsiella
Usually pathogenic Mucoid growth
Found in intestinal tract Polysacchride capsules
Facultive anaerobes but grows better aerobically Lack motility
Diffretiated on salmonella shigella agar Lysine Decarboxylase(+)
Selected by XLD, SS , HE agar. Citrate(+)
Entiched by Selenite F and tetrathionate broth Voges Parauskauer(+) Commented [DK4]: Selective?
May Causes bactermisa Present in the resptory tract and feces .
K.Pneumonae is known to cause pneumoniea and
haemorage in the lungs.
Causes UTI
Proteus
Found in the intestinal tract and cuases infections
only when it leaves the intestinal tract.
Found in urinary tract infections and cause
beacteremia and pneumonia in
immunocompromised.
Urease(+)(alkalizing and promote kidney stone
formation)

Bile esculin agar Streptococcal selective agar

Purpose: Encourage growth of lactic acid bacteria

Selective: Enriched
Selective Differential Inhibits commensal Constains sheeps blood
Bile salts select out Differentiate enteric bacteria , to cultivate
gram positive bacteria . enterococcus from contains neomysin sterptococcus
streptococcus. and polymyxin
Enterococcus
hydrolyze esculin to inhibit gram-negative
products that react with bacilli and
ferric acid to blacken staphylococci,
the medium
Selects for Group A
Essential Components: streptococci
Polysorbate 80 Provision of fatty acids needed
specifically by Lactobacilli
Sodium acetate and ammonium citrate
Prevention of Streptococci, moulds and many other Appears positive when beta clear zone haemolysis
microorganisms from colonisation on agar is observed
Magnesium sulfate and manganese sulfate
Provision of essential ions for Lactobacilli to
multiply.
Yeast extract Contains vitamins and amino
acids specifically needed by Lactobacilli.
Expected Positive/Negative Results:
Positive: Large, clear and white colonies
Negative: No colonies
Expected Positive result:
Group A Streptococci appear as translucent/ white
small colonies surrounded by a clear zone
Negative Result:
No formation of colonies/ No hemolysis of blood

Lactobacillus-MRS agar MRVP Broth

Selective
Contains sodium acetate which suppressed Voges Proskauer Voges Proskauer
other competing bacteria Positive Negative

Polysorbate 80, increases nutrient uptake by

laactobacillus Methyl Red Methyl Red
Positive Negative

XLD Agar HE Agar

 Selective: Differential :
Selective: Differential : Inhibits Gram Positive Diffrentiates Shigella
Inhibits Gram positive Diffrentiates Shigella Bacteria and selects for and salmonella
& enteric bacteria. and salmonella Gram negative bacteria (salmonella produces Commented [DK5]: Not shigella
Inhibits proteus Hydrogen sulfide,
Selects for Shigella colonies seen as black)
and salmonella
Contains sodiul thiosulfate and ferric ammonium
citrate
1. Shigella is non 1.Salmonella is xylose Salmonella appears as Shigella appears as
xylose and sucrose fermenting black colonies as they green colones
fermenting and . Acid causes phenol metabolise thiosulfate
no acid is red to turn orange and produce H2S
produce . It 2.Xylose is used up. *Proteus sp. and Citrobacter freundii, produce
metabolises Salmonella will H2S, but these organisms are usually inhibited by
lysine and will produce lysine the bile salts in the HE agar.
remain pink decarboxylase to
metabolise lysine and Brommenthol Blue
cause the agar to be Fermentation/salicin Green/Blue no
basic metabolism and acid fermentation and
3.Salmonella production turns agar Neutral/Basic
metabolises thiosulfate orange*
and produces Black
colonies. Only Non coliforms are able to use Commented [DK6]: Coliforms include Citrobacter
Sodium deoxycholate salicin/lactose/sucrose Enterobacter
Hafnia
Inhibits Gram Positive and other Enteric Bile Salts inhibit Gram positive bacteria
Klebsiella
Bacteria Escherichia
Novobiocin: Inhibits Proteus bacteria which can
metabolize the agar like Salmonella

Oxidase Test Indole Test ONPG Test

Test for aerobes which have determine the ability of the
Cytochrome c oxidase organism to convert
tryptophan into the indole

TSI Nitrate Reduction test

Lactose is Glucose is neither
fermented fermented and lactose/sucrose
and acid is lactose is not nor glucose is
pproduced fermented, both
the butt and the
slant will be
red.(Basic and
neutral)

H2S is produced
 Antibiotics

Antibiotic Resistance
Vectors for Antibiotic resistance usually Produce Proteins to
degrade or prevent the Antibiotics from killing the bacteria or
functioning
Name Class Mode of Action*

Kanamycin aminoglycoside Binds 30S ribosomal subunit; causes mis- Bactericidal

translation
Spectinomycin aminoglycoside Binds 30S ribosomal subunit; interrupts protein Bactericidal
synthesis

Streptomycin aminoglycoside Inhibits initiation of protein synthesis Bactericidal

Ampicillin beta-lactam Inhibits cell wall synthesis Bactericidal
Carbenicillin beta-lactam Inhibits cell wall synthesis Bactericidal
Bleomycin glycopeptide Induces DNA breaks Bactericidal
Erythromycin macrolide Blocks 50S ribosomal subunit; inhibits aminoacyl Bacteriostatic
translocation
Tetracycline tetracycline Binds 30S ribosomal subunit; inhibits protein Bacteriostatic
synthesis (elongation step)
Chloramphenic Binds 50S ribosomal subunit; inhibits peptidyl Bacteriostatic
ol translocation

Kirby Bauer Test

- Draw a - Place sterile filter Dip the swab in broth Use sterile forceps to
quadrant on paper disk in each containing place filter paper disk in
the back of the quadrant microorganism respective quadrant
petri dish and - Dispense 5l of - Entire agar Incubate for 16- 18 hours
agar plate and each antibiotic onto plate is
label with the disks and air dry swabbed
codes - Left to dry

Muller Hinton Agar

 Nonselective Non-Diffrential

Properties:
Contains starch : Protects against toxins Allows
growth of bacteria.
Loose Agar: Allows Diffusion of Antibiotics
Non Selective and Non Differential.

Depth and firmness of agar can affect antibiotic

diffusion
Too Thick or Firm agar reduces the ability of
the antibiotic to diffuse
Too thin or too loose agar allows the Antibiotic
to diffuse too fast

Thickness of the disc used can affect the test.

Amount of antibiotic can affect the effectiveness

of the antibiotic diffusion test
pH of 7.2-.4 should be maintained.

Synergistic effects are interaction or cooperation of Antagonistic if the activity of one drug is
two or more agents that produces an effect greater compromised in the presence of the other. => B
than the sum of their individual effects. => A

Minimum inhibitory concentration

Minimum inhibitory concentration
Serial Dillution
- Folic acid metabolism required to form nucleic acids , Adenine and Thymine .
- Antibiotics : Sulphonamides ( used in inflammatory bowel disease ) , Trimethoprim ( treatment of
bladder infection )
- Mechanism of action : Sulphonamides ( bacteriostatic )
- Competitive inhibitors
- Competes with para-aminobenzoic acid ( PABA ) with the enzyme dihydropteroate synthase
(DHPS)
- Prevents PABA from being added to folic acid molecule to form folate , essential for formation of
Adenine and Thymine
- W/o folate , not able to synthesise DNA and RNA for metabolic functions
- If Sulphonamides and Trimethoprim work hand in hand , require lower dosage of both and function
is stronger.
 Add from
rDNA ,

Broth dilution MIC

Disk diffusion Kirby Bauer

Micro vs macrodillution

PCR
Look though the
Serology

Ouchterlony
To determine if two antibody are related Commented [DK7]: Cfm with facil
Spurshttp://jeeves.mmg.uci.edu/immunology/Ouchterlony/Ouchterlony1.pdf

Complete Identity The lack of spur cross mean that there is

complete identity
The two antigens are the same
Non Identity The presence of two spurs indicate that the
HAS and HTF are unrelated
 Indicates that the antigens are unrelated and
have different epitopes
Partial Identity The presence of one Spur means that the
antigens are related but not identical
Radial Immunodiffusion

Diameter of ring measures IgG concentration in patients

serum
Measures the severity of the infection
The amount of antigen is proportional to the standard
curvs.

An antigen of different concentration are used to create a

standard curve.
An antigen is then tested and identified
Ask if antigen identity is known or tested using ab

Advantages Disadvantages

Test Tube Inexpensive Long reaction time

Easy to carry out Semi-quantitative
Easy setup No precipitate formed if
there is no equivalence
between antigen &
antibodies

Ouchterlony Inexpensive Long reaction time

Easy to carry out Semi-quantitative
Can test multiple patients serum No precipitate formed if
there is no equivalence
between antigen &
antibodies
 Immunofluorescence
Purpose: To detect unknown antigen with
known antibodies
Principle:
- used on biological samples
- The specificity of antibodies to the
antigen
When is this technique used ?
- Immunostaining. Makes use of
fluorophores to visualise antibodies
location.
If fluorescence is observed under microscope ,
the antigen can be identified which can be
traced back to which pathogen.
Dark Field Microscopy

Free of artifects
Light used idoes not reach the eyepiece
Only scattered light limmuniates the image
Strog lihght is used and may damage the sample

It also allows us to observe highly magnified

images of
unstained samples.
For the identification of Treponema pallidum,
exudates and
fluids from lesions are examined as a wet mount
using
dark-field microscopy.

FTA-ABS Rapid Plasma Reagin

FTA -ABS is used to domfirm ig one has syphilis , it
checks for antibodies specific for treponemea
pallidium
Cannot be used to distinguesd f one has recovered
Antibodies are washed from the slide of no results
are obtained.
Blood sample is collected.
Faslse negative can occur , if you had syphilis for
less than 3 mounths ,
False positive results can occur due to similar
antigens.
- Detects for non-specific antibodies
against Syphilis
- Can be seen with naked eye
- Easy to use as comes in kit forms
- Requires less time
- Cannot be tested on CBF
test does not look for antibodies against the
actual bacterium, Treponema pallidum, but
rather for antibodies against substances
released by cells when they are damaged by T.
pallidum.

Commented [DK8]: Sensitivity vs specificity

Latex Agglutination
 - Observed when a sample containing the specific antigen (or antibody) is mixed with an antibody
(or antigen) which is coated on the surface of latex particles (example latex beads).
- If sample has target protein, binding occurs

f
 Zone of Equivalance
The range of optimal concentrations, at which the
greatest amount of precipitation is generated, is called
the equivalent zone. Based upon this principle, the
presence of specific antigens in a mixture and the
relative concentration of antigens can be detected
using antibodies.

ELISA
1. Fast duration required for ELISA result:
~ 3 hours
2. Highly specific: detects only the desired target molecule
3. Highly sensitive:
detects low amounts (pg/mL) of target molecule
Direct ELISA
The primary antibody specific for the antigen is
tagged
Quick because only one antibody and fewer steps are
used.
Cross-reactivity of secondary antibody is eliminated.
Immunoreactivity of the primary antibody might be
affected by tag.
No flexibility in choice of primary antibody label from
one experiment to another.
Minimal signal amplification.
Indirect ELISA
Only one tagged secondary antibody is needed
for many primary antibodies, one substrate is
also needed
Maximum immunoreactivity of the primary
antibody is retained as it does not have enzyme.
Sensitivity is increased as many secondary
antibodies can bind to one primary antibody
Cross-reactivity might occur with the secondary
antibody, resulting in nonspecific signal.
An extra incubation step is required in the
procedure.

Direct Capture and

Direct detection
Direct Capture and
indirect detection
Indirect Capture
Direct detection
Indirect Capture
indirect detection
Anaerobic Bacteria

Anaerobic Bacteria Aerobes and facultative

Obligate Anaerobes: Require reduced oxygen
Do not posses tesnsion.
Catalase nor SOD.
Require reduced Do not posses
oxygen tesnsion. Cytochrome C for oxygen
metabolism
Low levels of SOD
Low levels of Catalase
aerobes
Possess Cytochrome C
detected by the oxidase
Test
Posses Catalase which
catalyses ROS hydrogen
Peroxide :
2H202 -> 2H20 +O2

SOD which catalyses

O-2 +O-2 +2H+ -> H2O +
O2

Thioglycollate broth
How does it differentiate?
Obligate aerobes grow in this oxygen-rich top layer.

Obligate anaerobes will only grow in the lower areas of the tube.

Microaerophiles will grow in a thin layer below the richly-oxygenated layer.

Facultative or aerotolerant anaerobes can grow throughout the medium but will primarily grow in the
middle of the tube, between the oxygen-rich and oxygen-free zones.

Colostrium Spp.
Anaerobic Bacilli
Spore(+) Oxidase(-) Catalase Gram(+) Naglers Reaction: Bacilli
(-) Positive

Colostrium tetani Clostridium Clostridium Clostridium

difficile
Germinate in tissue Causes
and pseudomembraneous
producesneurotoxins. colitis.
Suoresses action Part of the normal
potential which causes Gastrointestinal flora but
muscle spasms becomes pathogenic when
delocalized or proliferates
after antibiotics use..
Antibiotic resistant
botulinum perfringens
Spores found in - Obligate anaerobe
preserved canned food - Fastidious bacteria
Germinate in anaerobic - Causes Gas Gangree.
environment. - Accuminalation
of H2 and Co2
Produces neurotoxin. - Produces Alpha toxin or
MOT:Ingestion of Haemolytic
contaminated and Exotoxin(Lectinase)
inadequately prepared - MOT ,food route or
food. injuries and wounds

Produces Toxin A & B

Neisseria
Aerobic Diplococcus
Catalase+ H2S (-) Haemolysis(-) Gram (-) Coffee bean
Oxidase(+) No shaped diplococci
5% Co2 /coccus?
Ferments carbohydrates and produces acids.
Time taken to grow: 24 hours
Temperature: 35C
Requires blood or hemoglobin and several amino acids and vitamins in order to grow
Pili present, which play a major role in adherence, extend several micrometers from the cell surface
Media containing hemoglobin are needed for isolation and growth of the organism.
Medium: Chocolate agar (with carbon dioxide)

N. meningitidis
Semicircular diplococcus
Upper respiratory tract infections.
Sputum samples can be collected.
LPS, causes strong acute inflammatory response.
Causes bacterial meningitis, the causative agent of
meningococcal septicaemia.
Nasopharangeal Blood spcimmems and spinal fluid
samples may be obtained.
Opa proteins inhibit immunr cell receptors
N. gonorrhoeae
Oxidizes only glucose and grows slowly
Por Proteins prevent lysosome phagasome
function.
Has Pilli for host attachment
Changes antigenic structure by mutation
IgA protease
Found after sexual contact with an infected person
Causes gonorrhea
Causes Skin lesions papules and can also be
cultured from joint fluid

Mycobacterium .spp
Resistant to Gram staining
Stained by Acid Fast.

The Cell wall has high levels of mycolic acid making them
impermeable to gram stain, alcohol or acids.

Can live intracellularly

Mycolic acid causes bacteria to grow slowly as mycolic acid

has low permeability and causes clumping of bacteria causing
reduced space and nutrient uptake rate.

Tween 80 Oxidase(+) Tellurite(+) rod-shaped

hydrolysis test: Catalase(+) organism
Optimum growth temperatures: 30 C to 32 C (vary according to species)

Mycobacterium Tuberculosis Mycobacterium leprae

Why Fastidious? - Skin Scrappings/nasal sputum can be used
- Cord factor (Virulence): Prevents fusion - Form lesions
between phagosomal vesicles - Neurologic distrurbances
- Leprosy skin lesions progressive
Symptoms: Bloody cough, fever, scarring of lungs,
damage to rest of the body
fatigue, chest pain
- Can be tested by tuberculin skin test, by
injecting tuberculin and looking for swelling
within 2 days

Acid Fast Stain

Campylobacter spp. Legionella pneumophila
- S-shaped Catalse(+)
- Gram negative Oxidase(+)
- Has flagella (motile) Aerobic
- Commonly found in Poultry Fastidious as they grow luxuriantly on
tapwater.
- Both catalase and oxidase test positive.
It is caused by Legionella pneumophila,
Special Requirements it requires are
usually found in air-conditioners, spas,
- Most optimal at 42 C cooling towers etc. It is usually spread by
- Needs to be cultivated at low oxygen breathing in mist that contains the bacteria.
environment. Signs and symptoms:
Cough, shortness of breath, high fever,
muscle pains, and headaches,mental
changes
Specimen collected:
taking a sample of sputum (phlegm)
Treatment:
Antibiotics (eg: ciprofloxacin, levofloxacin)
Needs to be cultured on charcoal-containing
agar,requires L- cysteine and iron to grow.
35-37C in moist environment
Requires nutrients from tap water to grow.
- Live in water , but sperads thugh the
nasalpharanyal root.
- Requires iron
- Not stained by gra,
- Bcye agar

Helicobacter pylori Brucella spp.

 Morphology: Gram-negative, spirillum, Morphology:
microaerophilic bacterium Gram-negative
Requirements: Requires low oxygen conc. Coccobacilli
Virulence factor: adhesin, urease and Non-spore forming
flagella Non-motile
Culture: Brain Heart Infusion (BHI) Broth, Special Requirements:
Chocolate Agar Enriched media at 37 and pH 6.6 to 7.4
Aerobes

Bacteroides fragilis Haemophilus

influenzae
Infects respitory tract

Chlamydia
Intracellular obligate parasites as they cannot synthesize their own ATP
Live only in the human host
Spreads by direct contact
Trophic for cuboidial cells
Momps bind to host and undergoes endocytosis
Reticulate bodies form inclusions which are counted
Gram staining not used
Giemsa stain use
Causes Genital pulpuled, yellow discharge, pain nausea and precoucious
conjuvites.
RB Elementry bodies Giemsa
stained cells

1. Immunofluorescence 2. Rapid staining method 3. Giemsa stained 5. Cell culture

- Finds Chlamydia antigens
- Use fluorescent miUse
fluorescein-conjugated
antibodies
- croscope
- Chlamydia trachomatis
exhibits a brilliant apple-
green fluorescence
- Combination of methyl inclusion bodies
green + neutral red
stains, washing at pH
5.0, inclusions stained
red while cell cytoplasm
was pale pink and cell
nuclei were pale green
- After that, use dark field
microscopy

Chalmydia cell culture Highly specific and sensitive

(Development takes 1-2 day s) 1 Chlamydia are obligate intracellular parasites , they cannot produce
their own ATP and require Host ATPso cell culture is used

2 Infected Sample lysed by sonication or glass beads Sample

3 Hela cells or McCoy(Fibroblast) trophic for chalmydia is inoculated

with sample

4 Cycloheximide added ; reduces the cellular

metabolism of host cell, giving the bacteria more substrates ;
increase sensitivity by increasing the inclusion number
Polycation diethylaminoethyl-dextran added : shield charges on the
Eukaryotes , as both cells have a net negative charge on their
membrane, increases infectivity

Buffered Charcoal Yeast Agar Lowenstein Jensen Agar

Needs to be cultured on charcoal-containing Contains homogenized eggs and aminoacids
agar,requires L- cysteine and iron to grow. supports mycobacterium growth.n
35-37C in moist environment Selective Enriiched
Requires nutrients from tap water to grow. Contains Glycerol
Charcoal Yeast Extract (CYE) Selects for which encourages
Or mycobacterium Mycobacterium
Buffered Charcoal Yeast Extract ( BCYE) tuberculosis growth
for legionella Contains mechalite Or
green and congo red
which prevents growth Pyuverate which
of other bacteria favours M. bovis

Chocolate Agar Chocolate Agar w Factor V & X

Contains lysed red blood cells that have been Contains lysed red blood cells that have been
lysed at 80 deg lysed at 80 deg
Haemophilus species,
Enriched Enriched
Supportsfastidious resipitory Supportsfastidious resipitory
bacteria bacteria

Contains Factor V NAD

Factor X Hemin
Thayer-Martin agar (modified chocolate agar) Cystine-tellurite blood agar
Thayer-Martin agar (modified chocolate agar) Meat heart and Tryptose : necessary nutrients (
nitrogen and carbon compounds , AA and trace
Selective Enriched: minerals ) for growth of C. diphtheriae
Selects for Nessierua
Contains lysed sheeps Sheep blood :growth factors for C. diphtheriae
Antibiotic used blood. As Neisierria Sodium chloride : supplies essential electrolytes
Vancomycin - Inhibits cannot lyse RBC and maintain osmotic equilibrium
most gram positive L-cystine : source of AA , enhancing H2S
bacteria production
Colistin - Inhibits most Tellurite : inhibit most upper respiratory tract
gram negative bacteria normal flora and inhibit growth of Gram-(-)/( +),
except Neisseria Tellurite will be reduced to tellurium , producing
Nystatin - Inhibits grey to black colonies
yeasts Agar
Cystine Tellurite Agar Base is used for the
selective isolation and differentiation of
Corynebacterium diphtheriae .
Some strains of Staphylococcus and Streptococci
can also grow here

Cefsulodin-irgasan-novobiocin (CIN) agar Bordet-Gengou agar

Highly selective for the isolation of Yersinia Components:
enterocolitica. (Sodium deoxycholate, cefsulodin, Blood
irgasan, and novobiocin + antibiotics) Potato extract
Differentiate between mannitol fermenting and Glycerol
non-mannitol fermenting organism Antibiotics Cephalexin/Penicillin
Positive result: colonies with deep red center with Function:
a transparent margin (ferments mannitol Enriched medium
producing acid turn colonies red color Isolation of Bordetella pertussis
Yersinia enterocolitica)

Parasitology

Diseases by unicellular parasites (protozoa)

Blood and tissue-malaria (highest number of
parasiterelated
death), leishmaniasis (second highest number of
parasite-related death), African trypanosomiasis
Urogenital tract trichomaniasis
 Intestinal amoebiasis, giardiasis
Diseases by multicellular parasites (helminths)
Nematodes or roundworm loiasis, hookworm
infection
Trematodes or flukes schistosomiasis
Cestodes or tapeworm T. saginata infection
Example
Transport temperature:
Blood: Refrigerated
Slides (both thin and thick): Room temperature
Specimen stability:
Refrigerated: 72 hours.
Room temperature: 36 hours.
Rejection criteria:
Clotted, frozen, hemolyzed, overfilled or underfilled tubes; Slides not kept at
room
temperature.
Leishmaniasis
Leishmania spp.
Leishmaniaa are Dimorphic protozoa .

They disrupt the Toll Like receptor of

macrophages and prevents fusion of
lysosome with the vacuole

macrophages, skin
Sandfly bites
Caused by sandfly bite
Sympptoms:
Silent infection they cause skin infections
and fever and infect internal organs.
Can be diagnosed though a biopsy for liver
spleen and bone marrow/
Latext agglutination test is avaliablew

Trichomoniasis
Trichomonas vaginalis
Protozoa that only exsist as trophozoite
Has Adhesion factors for Vaginal epithelial cells

Intestinal, mucosal, genitourinary

Sexually transmitted with infected individual
Crvical smear can be used but has low sensitivitry
Cervical discharge sample is obtained.

African trypanosomiasis
 Trypanosoma brucei (African sleeping
disease)

Blood and tissue

Tsetse Fly bites

Loa loa filariasis

Loa Loa is a type of
helmith or roundwoem,
it can invade the
testes,lngs,kidney,heart
,eyes and nose.
Migrate in blood in day
and to vascular parts of
lung at night

Eye
Horsefly bites

Symptoms include swelling, joint pain, pan in eye,

fatigue and itching,and muscle pain,
Mimics the host antigen, induce T cell anergy and
evades complement system

Can be diagnosed with a blood specimim on glass

s;ode
Hookworm infection

Caused by Ancylostoma duodenale

Small intestine/GI tract

Walking barefoot on contaminated soil
Proudces Hyaduoridase and has buccal capsule.
Symptoms: Anaemis, Coughing, Bloody stool, lost
of appetite and abdominal pain
Treatment:
Iron supplement
Albandozoe
Mabendozole
Stool sample is collected

Plasmodium
i) Plasmodium falciparum
ii) P. vivax,
iii) P. ovale
iv) P. malariae

) From the diagram above, describe how the malarial parasite is transmitted to humans?
Biten by an infected female mosquito of genus Anopheles, which previously had stung a person infected
with malaria.

b) Name the etiological agents of malaria

i) Plasmodium falciparum
ii) P. vivax,
iii) P. ovale
iv) P. malariae

c) Out of these etiological agents, which is the most deadly? Why is this so?
 Plasmodium Falciparum. It multiplies rapidly in the blood thus able to form anaemia. It can also clog
blood vessel.

d) The life-cycle of the Plasmodium parasite is complicated as it has many stages. What are the
various stages?
1. infection of a human with sporozoites
2. asexual reproduction
3. sexual reproduction

Sporozoites---Enter the tissue and circulate to the Liver---> Schizont

Schizont ---> Contains many Merozoites. ---> Mezoiutes bud and released to circulation.
Merozoites infect the erythrocytes . Trophozoite are formed and grow , replicate the rbc.

e) What happens during each stage of the Plasmodium parasites life cycle?
1. infected female anopheles mosquito bites a person and injects sporozoites saliva into blood
circulation

2. sporozoites enter the liver and start dividing leading to schizonts creation in 6-7 days. each schizont
give birth to thousands of merozoites and then released into bloodstream.

3. parasites begin to develop into sexual form of parasite aka gametocyte, which is required for
productive transmission to the mosquito host. Female and male gametocytes are taken up by mosquito and
both gametocytes undergo sexual phase of life cycle resulting in zygote then form oocyte. when the oocyst
rupture, haploid sporozoites are released, they migrate to salivary glands and transmission to humans
happens.

f) Through your understanding on the life-cycle of the Plasmodium parasite, decide on a good
specimen to use for diagnosis.
Blood cell, with giemsa or stain to see the parasite in the cell
 g) Likewise, how do you use the knowledge regarding the life-cycle and/or modes of transmission of
other parasites to decide on a good specimen for diagnosis?
As malaria is transmitted in blood stream, the specimen for malaria is whole blood.

The first step is to centrifuge blood first, followed by an incubation time of 9 days to 30 days depending
on species. It take 1 hour for the sporozoites to enter the liver and a generation time of 48 hours in
the erythrocytes.

Mechanism Example
Mechanical damage
Physiological disturbance
Tissue destruction Hookworms
Inflammation Trypanosomas
trypomastigotes
and
Mechanical damage in tissue;
Immunopathology
 Loa loa filariasis
Loa Loa

Eye
Horsefly bites

1. Once a patients specimen is obtained, there are many diagnostic methods that can be used to diagnose
for the etiological agent for malaria. List and describe each diagnostic method, and explain the rationale
behind each method.

Diagnostic method Rationale behind method

1 Thick blood film Able to detect the presence of plasmodium as it uses a larger sample of
Place a drop of whole blood thus, is more sensitive than thin blood film. However, it is unable to
blood onto slide allow recognition of the species of the plasmodium.
Allow to dry
Stain with Giemsa stain
or Wrights stain
Observe under a light
microscope at 1000X
magnification
2 Thin blood film Allow us to detect parasite and abnormal cells in the blood. This includes for
plasmodium. It also allow us to observe the morphology of the plasmodium
thus able to know the species .

3 Fluorescence Uses a high intensity light source which excites a fluorescent species in a
microscopy sample of interest.

4 Molecular method: Ability to detect for low levels of parasites in the blood (5 parasites or less/ ul
of blood) and allow identification of parasites to species level

(Nested and multiplex PCR methods can give valuable info when difficult
morphological problems arise during attempts to identify parasites to the
species level. PCR based methods are also useful for studies on strain
 variation, mutations and studies of parasite genes involved in drug
resistance)
5 Other tests: Serological tests.

2. Each diagnostic method has its pros and cons. Discuss the challenges faced while using these
diagnostic methods for the diagnosis of malarial parasite.

Diagnostic Challenges
method
Blood films Thick films: unable to see the morphology of the plasmodium thus unable to
recognize the species

Thin films: less sensitive as not much blood sample is examined

Fluorescence QBC: more technically demanding, requires specialized equipment to separate cell
microscopy layers by centrifugation and good fluorescence microscope with high-intensity
mercury vapor / quartz halogen lamp to provide excitation wavelength

AO: Nonspecific & stains nucleic acids from all cell types; microscopist must learn to
distinguish fluorescence-stained parasites from other cells & cellular debris
containing nucleic acids; not specific for malaria parasites but specific for P.
falciparum
PCR can give false positive results, not quantitative, performance is easily affected by
contamination.

3. Can these methods be used to diagnose for other parasitic diseases too? Give examples.

Yes, blood film method can be use to diagnose filariasis.

Diagnostic Challenges
method
Blood films Thick films: unable to see the morphology of the plasmodium thus unable to
recognize the species

Thin films: less sensitive as not much blood sample is examined

Fluorescence QBC: more technically demanding, requires specialized equipment to separate cell
microscopy layers by centrifugation and good fluorescence microscope with high-intensity
mercury vapor / quartz halogen lamp to provide excitation wavelength

AO: Nonspecific & stains nucleic acids from all cell types; microscopist must learn to
distinguish fluorescence-stained parasites from other cells & cellular debris
containing nucleic acids; not specific for malaria parasites but specific for P.
falciparum
PCR can give false positive results, not quantitative, performance is easily affected by
contamination.
Trypanosomas

Lifecycle and reproduction(Tufail)

Signs and symptoms(Jezz)
Pathogenic factor and characteristic(Tessa) .,
Diagnosis(Darius)
Treatment(Jacelyn)
Diagnosis
Lymph Node Aspirate
Giemsa stained parasites affords high specificity to thetrained eye
o Low sensitivity(20000 /ml is needed)
o motility and morphology can be used to identify tryposomas
o Cheap and time consuming
o High Sensitivity(500 per mL) can be archived by centrifuging, tryposomas would be
concentrated. Tryposomas are stained with arcmine orange , and will be found as
flurescent in the buffy coat
o

CATT Test (Serological)

o Antibody against LiTat 1.3. Antigen is detected
o Positive when antibody antigen crosslink
o Succeptible to false-negative when the pathogen does not have LiTAT1.3
o Succeptible to prozone effect where too much of wither antige or antibiody

o
Clinical Presentation and pathology
increased immunoglobulin concentrations, especially of IgM . Low serum C3 levels

6y
Treatments
Stage One
Intramuscular or intravenous pentamidine

Stage Two
Intravenous injection of combination of nifurtimox and eflornithine (NECT) or eflornithine
- eflornithine alone appear to be more effective and result in fewer side effects.

Life cycle.
Infective metacyclic trypomastigotes deposited into thru salivary gland human as the tse2 fly take in blood.

metacyclic trypomastigotes transform to bloodstream trypomastigote of a long and slender form

it travels thru the bloodstream and lymph. spinal fluid. They also replicate by binary fission at this point. .forming short
and stumpy.it is extra cellular

which then the tse2 fly take in when it wants to take in blood.

the bloodstream trypomastigotes(SHORT STUMPY) transform to procyclic trypamastigote and carry on replicating

when it reaches the midgut, it transform to epimastigote.

it then goes to salivary gland and continue replicating

it transform into metacyclic trypomastigote before deposting to mammal

THEY CHANGE MORPHOLOGY as to adapw the environment such as pH and nutrient avail.

Virology
 Non enveloped viruses:
1. The outermost covering is the capsid made up of
proteins
2. Non enveloped viruses are more virulent and causes
host cell lysis
3. These viruses are resistant to heat, acids, and drying
4. It can survive inside gastrointestinal tract
5. It can retain its infectivity even after drying
6. It will induce antibody production in the host
7. Mode of transmission is through fecal or oral matter,
formites and dust
DNA viruses
Location of genome replication Nucleus
Location of viral protein synthesis Ribosome
Enveloped viruses
1. The outermost envelope is made up of phospholipids,
proteins or
glycoprotein which surround the capsid
2. Enveloped viruses are less virulent often released by
budding and
rarely cause host cell lysis
3. Are sensitive to heat, acids, and drying
4. Generally it cannot survive inside gastrointestinal tract
5. It lose its infectivity on drying
6. It will induce both cell mediated and antibody mediated
immune
response in the host
7. Mode of transmission is through blood or organ
transplants or through secretions
Characteristics of viral envelope
Made of lipid and proteins rarely glycoprotein
May be modified host plasma membrane or internal
membranes
Projections from the envelope are known as spikes or
peplomers
RNA viruses
cytoplasm
Ribosome
 Type of
Example of
Virus Family nucleic Disease caused by virus Portal of entry
virus
acid
Variola major
Via inhalation of
Poxviridae and Variola dsDNA smallpox
minor respiratory secretions
Parvoviridae inhalation of droplets
Parvovirus
ssDNA B19 infection through respiratory
B19
tract
Mouth (Fecal-oral
Picornaviridae poliovirus ssRNA (+) Polio
route)
respiratory inhalation of droplets
Paramyxoviridae syncytial ssRNA (-) Measles through respiratory
virus tract

ssRNA-
Retroviridae HIV AIDS Sexual intercourse
RT(+)

Hepadnaviridae Partial Sexual intercourse,

Hepatitis B Hepatitis B
dsDNA intravenously
Usually not bad at all but can Cancer free(reo have
cause diarrhea in children and fetistic tropoism for
Reoviridae Rotavirus
dsRNA immunocompromised ras actuve cancerous
Coeliac in immunodeficient cells
 Class I -
Double-stranded
DNA (Smallpox)

Class II - Single-
stranded DNA
(Adeno associated
Virus)

Class III - Double-

stranded RNA
(Reovirus)

Class IV- Single-

stranded (+) sense
RNA (Dengue
Virus)

Class V - Single-
stranded (-) sense
RNA
(Influenza virus)

Class VI- Single-

stranded (+) sense
RNA with DNA
intermediate
(Retrovirus)
Class VII Double-
stranded DNA with
RNA
intermediate
(Hepadnaviruses)
1.Attachement The virus attaches itself to the receptors found around the host cell

Viruses with no viral envelope enter Viruses with a viral envelope mostly
the cell through endocytosis; they are undergo membrane fusion
ingested by the host cell through the
cell membrane

2.Pentration Virus enters the cells through endocytosis

3.Uncoating The envelope of virus is opened and genetic materials (DNA/RNA)

is freed into cytoplasm (DNA into the nucleus of the host cell)

4.Translation Protein building machinery of host cell translates the instructions

into the components of a new virus

5. Assembly The parts are then assembled into new viruses within the host cell

6.Release -New viruses emerge are released by lysis of the host cell
-Usually killing the host cell in the process
Most naked virus undergo lysis
 Most enveloped virus undergo budding

Drug Mechanism of Action Viral Spectrum

interference with the viral protein, M2, a proton channel// Blocks M2

Amantadine protein channel (type A only), disrupts hydrogen transport, viral
uncoating in host cell and therefore viral RNA transcription. Influenza A
have cidofovir diphosphate that inhibits viral replication by
selectively inhibiting viral DNA polymerase. incorporates
Cidofovir itself into viral DNA hence inhibiting viral DNA synthesis //
Inhibits viral replication by selectively inhibiting DNA Herpesvirus
polymerase. Insert itself into viral DNA hence reducing rate
of DNA synthesis.
(Ours)
Didanosine
HIV-1, HIV- 2

(Ours)
Darunavir HIV protease inhibitor
HIV 1, HIV -2
interfering with the synthesis of viral DNA herpes simplex and
Vidarabine Viral Polymerase inhibitor: interfering with synthesis of viral varicella zoster
DNA viruses.

Neuraminidase Inhibits neuraminidase and virus replication

Influenza A and B
inhibitor

Epidemology
Colony forming units
Applied to:
Spead plate
Colony forming unit
per ml (CFU/ml)
Number of CFU
on each plates are
counted.
Select plates with
manageable
numbers of
colonies
(30 to 300
colonies).
Plates with > 300
colonies: TNTC (too
numerous to count)
Plates with < 30
colonies: TFTC (too few
to count)
Morphology
Bacteria Shapes
 Cocobacillus

Vibrio

Coccus

Spirillum

Spirochete

Bacillus