P SYTEMIC BACTERIOLOGY LAB DIAGNOSIS

a Gram positive cocci Speci Micros Isolation and Bio Serological test
g men copic examination identification from chemical test
e culture
1.S. aureus Wound swab, Gram staining Culture in: a)Catalase test: A.There are no generally useful
pus, sputum, findings: i)Blood agar media positive serologic or skin test.
| blood, CSF. They are arranged in ii)Nutrient agar media b)Coagulase test: B.Detection of anti-teichoic antibody
grape-like irregular iii)5%-10% NaCl Positive may be done.
clusters. Finding: Golden iii)Mannitol
1 yellow colored colony. fermentation
test:positive
iv)DNAase
test:Positive
2.Strep. pyogenes Throat swab, Gram staining Culture in: a)Catalase test: A.ASO titre
pus, sputum, findings: i)Blood agar media negative B.anti DPNase
blood, serum. They are arranged in Findings: Pin point b)Coagulase test: C.Anti-DNAase-B
chain colony surrounded by negative D.Anti-hyaluronidase
a clear zone of E.Anti-streptokinase antibody
hemolysis
ii)Culture also in Mac
Conkey’s agar media.
3.Strep. Sputum, Gram staining i)Blood agar media a)Inulin fermentation Capsular swelling test (Quellung
pneumoniae blood, CSF. findings: Findings: Alpha test reaction)
Gram positive hemolytic colony. b)Optochin sensitivity
diplococci, lanced ii)Culture also in Mac test
shaped. Conkey’s agar media. c)Bile solubility test

Gram Specimen Microscopic Isolation Biochemical Serological Others

negative examination and test test
cocci identification
from culture

N. Male: Urethral Gram i)Chololate a)Oxidase A. Nucleic acid based technique:

gonorrhoea discharge after per staining agar media positive ELISA PCR
rectal massage or findings: ii)Modified b)Ferments B.
morning drop of Gram positive Theyar Media glucose DNA probe
secretion diplococci and (Incubation assay
Female: pus cell. temperature:
Endocervical 37℃ ,
swab Incubation
period: 18-24
hours,
Incubation
Environment:
5-10% CO 2)

N. CSF and blood, Gram i)Chololate a)Oxidase Detection of Examination of CSF:

menigitidis nasopharyngeal staining: agar media positive bacterial Turbid due to pus cell
swab, joint fliud, Pus cell, ii)Mac b)Acid antigens from
skin lesion, intracellular Conkey’s agar production CSF by latex
trachial aspirate. Gram negative media from glucose agglutination
diplococci and maltose. test or other
test.
 Gram positive Specimen Microscopic Isolation and Biochemical test Other test
P bacilli examination identification from
a culture
g C. diptheriae 2 throat swab, Gram staining: i)Loeffer’s serum In vivo test: Schick NA based tech.:
nasal swab Gram positive bacilli- slope test PCR
e Chinese letter ii)Blood tellurite agar In vitro test:
Appearance Finding: a)Elek test
Albert staining: Black color of b)Tissue culture test
| Bacilli take bluish green tellurium in the colony
color & metachromatic
2 granules take brick red
color.
Cl. tetani Wound swab Gram staining: Robertson’s cooked A.
Gram positive bacilli meat media Aggluti
(tennis Finding: nation
Racket appearance) Turbidity of medium test
& blackening of meat. B.Toxi
Genicity
Test
C.
Animal
inocul
ation
test
Cl. Left over food Isolation from left Toxin identify
botulinum and patient’s over food. cation by:
serum Animal
patho
genicity test
Cl. Wound Gram staining: i) Robertson’s cooked Sugar fermentation Nagler’s
perfringes Swab, pus, Gram positive bacilli meat media Test (lactic acid reacion
Tissues from the ii) production)
wound Thioglycolate
Media
iii)Egg yolk test
B. Sputum, blood *stained by Mc.Fadyean i)Nutrient agar Serology:
anthrasis reaction ii)Blood agar (aerobic A.In vivo:
*Reaction is colonies are formed) neutra
positive(large, Gram lizing
positive rods in chains) test
B.In vitro:
Gel diffusion test.

Gram Speci Micros Isolation and Serological test Other test

negative men copic identification from
bacilli examination culture
E. coli Freshly passed Gram staining: i)Mac Conkey’s Sometimes helpful. For ETEC, EPEC,
stool, Finding: Gram agar (incubate at VTEC done in reference lab.
Rectal swab negative rods. 37 ℃ .
Finding: Form pink
colonies
Salmonella Blood for Not helpful i)Blood culture Positive from 2nd week.
culture(1st and (positive in 1st (Widal test)
2nd week), week)-done in 3
Serum for processes-
Widal test (2nd Traditional, lytic
week), Stool method, automated.
for culture(3rd ii)Stool culture-
week), Urine (positive from 2nd
for culture(4th or 3rd week).Done
week) in Mac Conkey’s
agar, DCA,
Shigella-salmonella
agar medium.
iii)Urine culture-
(positive from 2nd,
3rd and 4th
weeks).Done in-
Mac Conkey’s agar,
DCA.
 Shigella Fresh stool, Findings: Culture done in- a)Co-agglutination test Routine stool examination:
P rectal swab. *Plenty of pus Mac Conkey’s agar to detect the endotoxin A.
a cells or DCA or b)Slide agglutination test Physical-Odourless.
*Macrophage Salmonella- for confirmation of Mucus&
g *Some RBC Shigella agar Shigella and for blood in stool.
e medium determination of its
group.
B.
| Chemical: Alkaline stool
V. cholerae Stool, vomit, *Gram staining- i)Monsur’s a)Slide agglutination test Biochemical test:
3 rectal swab. Gram negative media-Small black b)Indirect Oxidase positive
comma shaped colonies surrounded haemagglutination test
bacilli. by characteristic c)ELISA
*Hanging drop hollows.
preparation: ii)Thiosulphate-
Typical motility Citrate-Bile-
(a fish in stream Sucrose agar:
appearance) Large yellow
*Dark ground colored colony
illumination:
Shooting star in
dark sky
appearance.
H. pylori Biopsy Non-invasive tests Invasive test
specimen of
gastric
a)Urea breath test i)Endoscopy & biopsy
mucosa,
b)Serological test:Ab detection ii)Rapid urease test
Blood for
serum c)13C bicarbonate assay iii)Histopathology & microscopy: Gram staining & Giemsa staining
antibodies. d)Antigen detection in urine & stool iv)Culture (Skirrow’s media at 42℃ )
e)PCR in stool, dental plaque and water v)PCR in gastric juice.
supplies.

Pseudo- Pus from Gram stain: Gram negative Culture:

monus The lesion. Rods. i)Grows in Mac Conkey’s agar medium
ii)Blue green pigment on ordinary nutrient agar and fruity aroma.
iii)Oxidase positive

Acid Fast bacilli Speci Micros Isolation and Immuno Other test
men copic exami identification from logical test/
nation culture Serological
test
M. tuberculosis 3 sputum *Ziehl-Neelsen i)Lowenstein Jensen i) Nucleic acid based tech
sample (now-a- staining-Findings- medium-Incubation Tuberculin nique-PCR
days 2 sample), M. tuberculosis at 37℃ for 6-8 test
bronchial appears as straight weeks aerobically. ii)ICT, ELISA
aspirates, or slight curved ii)Dubo’s solid for Ab for cell
blood, swab. rods. medium. mediated
*Dark ground iii)Middle brook immunity.
illumination- medium
Finding-Yellow-
orange bacilli.
*Auramine/
Rhodamine stain
(For rapid screening
purpose)
M. leprae Slit skin smear *Ziehl Neelsen No role No role A.
from 1 or 2 staining with 5% Animal inoculation test-In mouse foot-pad
sites from the sulphuric acid. in the armadillo.
 edge of leprosy *Smear shows acid B.
P lesion and fast bacilli within Lepromin test-The test is positive in T.
a nasal mucosa, macrophage. leprosy.
Scrapping from *The living cells are C.
g skin lesion or uniformly stained NA base technique:
e nasal mucosa. while dead cells PCR
fragmented and
irregular.
|

Laboratory diagnosis of syphilis

4
Specimen: Serous fluid the lesions of primary syphilis and secondary syphilis, Serum, CSF.

Microscopical examination:

Dark ground illumination: Cork screw movement of T. pallidum.

Or,

Direct fluorescent antibody test: Typical fluorescent stained spirochetes.

Isolation and identification from culture:

No role.

Serological test:

i)No-treponemal tests:

VDRL, RPR(Rapid plasma Reasin)

ii)Treponemal detection:

TPHA, FTA-ABS, ELISA.

Cocco- Specimen Microscopic Isolation and Serological test Other test

bacilli examination identification from
culture
H. influenzae Blood, CSF, Gram stained Chocolate agar media a)Latex A.Defini
naso- smear: Reveals & heated at 37℃ . agglutination test. tive
pharyngeal swab, coccobacilli -Pleomorphic growth b)CIE identif
pus is observed. ication by: Quellung reaction
B.NA based
Techni
que:PCR
H. ducreyi Tissue from the Gram stained Heated blood agar a) NA based
ulcer, pus smear: Reveals media supplemented Detection of H. Techni
aspirated from a coccobacilli with X factor. ducreyi to antigen que:PCR
lymph node. -Pleomorphic growth b)CIE
is observed

Obligate Specimen Microscopic Isolation and Serological test/ Other test

intracellular/ examination identification from Immuno
Wall-less culture Logical test
organism
Chlamydia Conjunctival swab, Cytology: Giemsa Culture: a)Antigen NA based
lymph node biopsy, staining Cell line culture detection: Techni
sputum, blood. (HeLA-229, BHK- Direct immuno que:PCR
21, Buffalo green fluorescence test,
monkey kidney ELISA.
cell) b)Serology:
High titre IgM
antibodies is
indicative.
Mycoplasma Throat swab, No role. Heart infusion a)Non-specific:
sputum, peptone broth. Cold agglutination
inflammatory titer of 1: 128 or
mediators, higher is indicative
respiratory of recent infection.
exudates, Urethral b)Specific:
or genital secretion. CFT: 4 fold or
greater rise in
specific antibody
titer.
 HI test
P IHT test.
a
g
e