By: ASM

Specimen:
Urethral discharge from men.
Urethral discharge and cervical secretions from
women.
A. Acute stages Lab. Diagnosis:
Smear:
Specimen can be stained by Gm stain and methylene
blue.
Gm staining Gm-ve diplococci IC in
polymorphnuclear leucocytes with few EC
organisms

Polymorphnuclear leucocytes

N.Gonorrhea EC

Culture:
On chocolate agar in CO2 enriched aerobic medium at
37C for 48 hrs.

Examination of the colonies can be identified by:

1. Film

Difference between smear and film:

The smear is an examination under the microscope
directly from the specimen, so well find the
polymorphnuclear leucocytes and the organisms are
IC and EC.
While the film is and examination under microscope of
the colonies taken after culture, i.e. no leucocytes
are found.

2. Oxidase test
It was supposed to
be on chocolate
blood agar of
N.gonorrhea,
but itll give the
same results

Other tests to confirm the diagnosis:

1. Production of acid from glucose.
2. Coagglutination test.
3. Antigen detection by ELISA.
4. Nucleic acid probe.

How can we diagnose N.Gonorrhea in the chronic

stage?
Specimen:
- Morning urethral drop or prostatic secretion in male.
- Swab from the cervix uteri in female.
Culture:
Selective culture medium: the Thayer and Martin
medium, which contains the antibiotics vancomycin,
colistin and nystatin (chocolate agar + VCN).
How can you treat an infection by N.gonorrhea?
Broad spectrum 3rd generation cephalosporins eg.
Ceftriaxone and fluoroquinolones eg. ciptofloxacin

Methods used for demonstration of T.pallidum:

1. Dark ground microscopy typical morphology and
motility.

Serological tests for T.pallidum:

Rapid plasma reagin test (PRP):

Other methods for demonstration of T.pallidum?

Staining by immunofluorescence staining technique.
Types of serological tests?
1. Cardiolipin Ab tests:
a. Venereal disease research lab. Test.
b. Rapid plasma reagin tests (PRP).
2. Treponemal Ab tests:
a. Fluorescent treponemal Ab test-absorbed.
b. T.pallidum particle agglutination test.

Type of specimen taken in:

1. Primary stage exudet from chancre.
2. Secondary stage exudet from skin lesions ,mucous
patches or condyloma lata.
3. Latent stage serum or CSF.
Tests done in each stage?
1. Primary stage dark ground microscopy,
immunofluorescence microscopy, nucleic acid and
PCR.
Cardiolipin Ab tests : -ve (early)
Fluorescent treponemal Ab tests: +ve
2. Secondary stage both tests are +ve
3. Latent stage-ve in serum and +ve in CSF

How can you treat a case of syphilis?

Benzathine penicillin or erythromycin or tetracycline or
cephalosporins

Cytological examination reveals inclusion bodies

What other lab. Diagnosis of chlamydia?

1. Isolation on cyclohexaminde treated MacCoys cells.
2. Direct IF
3. Nucleic acid probe, PCR and ELISA.
4. Serological tests: CF and micro IF.
How can you treat a case of chlamydial infection?
Tetracycline and erythromycine.
Topical and systemic sulphonamides.

By isolation and see the colonies and the microscope

How can you take the specimen?

-Urethral swabs or urine after prostatic massage in
males.
-Fetal membrane swabs and semen as a part of
investigating infertility.
Mention the types of mycoplasma causing sexually
transmitted diseases?
1. Mycoplasma hominis : female.
2. Ureaplasma urealyticum: male.
How can you treat a case of mycoplasmal infection?
tetracyclin

Sterile container
Sterile syringe

Sterile swab

By: ASM

1. Urine analysis : casts, blood, pus and bacteria.

2. Culture: blood agar, MacConkeys agar at 37 C
3.
a.
b.
c.

for 24 hrs, then antibiotic sensitivity test.

Colony count: to differentiate between true
infection and contamination by normal flora.
Calibrated loop.
Drop slide
Miles and Misra technique.

Semiquantitative methods
Calibrated loop

Surface contain
blood agar
Surface contain
MacConkeys agar
Urine is put here
Disadvantage: cant be used in mixed infection

Quantitative method

Separated colonies that can be counted

2 parameters to diagnose UTI:

a. Pus cells.
b. Bacteremia >10X5.

urinary TB
Example of the questions that may
come in our exam regarding the
following photos:
1. For what can we use LJ medium?
Its a selective medium for TB.
2. What are the decolorizing agents
used?
20% H2SO4 and 95% alcohol.
Enriched medium

selective medium

E.Coli

E.coli, klebsiella, proteus, pseudomonas

Lactose fermenting colonies

1. What are the biochemical reaction of the organism?

Indole +ve, citrate ve and methyl red +ve, ferment lactose with
acid production at 44 C (Eijkman test).
2. Enumerate the antigenic structure?
O, H and K
3. What are the virulence factors of this organism?
Type 1 fimbriae and and hemolysin.
4. Diseases produced by this organisms?
Cystitis, pyelitis and pyelonephritis, appendicular abscess,
peritonitis, cholecystis, septic wound, bedsores, bacteremia,
nosocomial infection and neonatal meningitis

Mucoid glistening colonies due to slime produced

On MacConkeys agar (lactose fermenter)

1. What are the biochemical reaction of the organism?

Indole -ve, citrate +ve and methyl red -ve, weak urease +ve.
2. Enumerate the antigenic structure?
Capsular antigens by capsular swelling test in wet films.
34. Diseases produced by this organisms?
K. Pneumoniae 3% of bacterial pneumonia, urinary tract
infection and hospital acquired infection.
K. Ozaenae ozaena.
K. Rhinoscleromatis rhinosclerosis.

E.coli

klebsiella

How to inhibit the swarming

effect?
The culture media should
inhibit the swarm by:
1. Blood agar contain 5-6%
agar.
2. MacConkeys or DCA.
3. CLED medium.
4. Contain inhibitory medium eg.
Phenyl ethyl alcohol or P-nitrophenyl glycerol

Non identical with line of separation

between them

identical and the 2 colonies

coalesce together

1. What are the biochemical reaction of the organism?

Strong urease +ve and can deaminate phenyl alanine by
deaminase to phenyl pyruvate.
2. Enumerate the antigenic structure?
O and H and bacitriocin typing for epidemiological purposes.
3. Diseases produced by this organisms?
1. Pr. Vulgaris nosocomial pathogen.
2. Bacteraemia, pneumonia or meningitis.
3. Focal lesion in debilitated patients.
4. UTI.

Its characterized by pigment formation: green, blue, red or black

1. What are the biochemical reaction of the organism?

Doesnt ferment sugar but oxidase G, oxidase +ve.
2. Enumerate the antigenic structure?
O and H and bacitriocin or phage typing for epidemiological
purposes.
3. What are the criteria for identification of pseudomonas?
Oxidase +ve, pigments and growth at 42 C
4. Diseases produced by this organisms?
Wound and burn infection, meningitis, UTI, RTI, otitis externa. Eye
infection and fatal septicemia.
5. How can you treat an infection by pseudomonas?
Combination of imipenum and ceftazemide.

It can resist heating at 60 C for 30 min

1. What are the biochemical reaction of the organism?

ferment mannitol with gas production.
2. What is the morphology?
Gm +ve cocci arranged in short chains or pairs, non motile and
non encapsulated.
4. Diseases produced by this organisms?
UYI, endocarditis, biliary tract infection, ear infection and
suppurative abdominal lesion.

Coagulase test
+ve
Staph. Aureus

-ve

novobiocin test
R
S
saprophyticus epidermidis

Non motile organism eg. klebsiella

Or enterococci

Highly motile organism eg. Proteus

Also it may be any other motile
organism eg. E.coli, pseudomonas

Nitrate reduction +ve

Nitrate reduction -ve

Oxidase +ve in pseudomonas

and may be also N.Gonorrhea

In enterobacteriacea
eg. E.coli, klebsiella, proteus

In non enterobacteriacea
eg. pseudomonas

All other bacteria

Proteus and may be

klebsiella (weak)