Laboratory Diagnosis

Syphilis Lymphogranuloma venereum (LGV) Chancroid Granuloma Inguinale (GI) Herpes Simplex

Aetiology

Treponema pallidium

Laboratory Diagnosis

Direct demonstration of organism in lesions Serological tests:

Non specific Treponemal test

Specific Treponemal Tests Flourescent Treponemal Antibody (FTA abs) - used to confirm a positive VDRL Test.

Veneral Disease Research Laboratory (VDRL) tests Rapid Plasma Reagin (RPR)

Specimens: Fluid/scraping from chancres (primary syphilis) Blood (primary and secondary syphilis) CSF (tertiary or neurosyphilis)

Specimen: Fluid/scraping from chancres

Dark field Microscopy Silver stain

Dark field Microscopy

Silver stain

- A nontreponemal serological screening test for syphilis.

Significance of VDRL test:
VDRL test becomes positive 1-2 weeks after appearance of (primary lesion) chancre. The test becomes reactive (50-75%) in the late phase of primary syphilis, becomes highly reactive (100%) in the secondary syphilis and reactivity decreases (75%) thereafter. VDRL test is also helpful in the diagnosis of congenital syphilis.

Since the test employs a non-treponemal antigen, false positivity occurs.

False positivity- biological false positives (BFP).
e.g. Pregnancy, malaria, infectious mononucleosis, hepatitis, relapsing fever, tropical eosinophilia, lepromatous leprosy, SLE, rheumatoid, arthritis etc. A reactive VDRL test does not necessarily imply that the person is syphilitic. Any reactive VDRL test must be confirmed with a specific or treponemal test such as FTA-ABS test, TPHA

Patient sera is diluted and non treponemal antigen added. Various degrees of clumping will determine the reactivity of the serum.
Non Reactive Weakly Reactive Reactive

Patient serum is treated to adsorb antibodies to any treponemes other than T. pallidum. The serum is then placed on a glass slide which was previously coated with Treponema pallidum ( grown in the testes of a rabbit). Anti-human globulin labelled with fluorescein dye is then added to the slide. Any antibodies to T. pallidum in patients serum will give a positive reaction. Positive FTA abs for syphilis

Aetiology

Haemophilus ducreyi

Specimen: Swab from ulcer base

Laboratory Diagnosis
Gram stain Culture on special media

Small gram-negative bacilli, intraand extracellularly dispersed, with "school of fish" distribution (arrows)

Aetiology

Calymmatobacterium (Klebsiella) granulomatis

Specimen: Scraping or biopsy material from ulcer

Laboratory Diagnosis:
Impression smear is made from a small piece of clean granulation tissue from the lesion on a glass slide. Giemsa or Wrights stain is done and slide examined for Donovan bodies

Large monocyte filled with Donovan bodies (Pund cell). Donovan bodies typically appear as encapsulated closed safety-pin.

Aetiology Chlamydia trachomatis Serotype L1, L2, L3

Laboratory Diagnosis: Specimen Pus from bubo Serum

Microscopy Stained smear (Giemsa) Electron Microscopy (EM) Immunofluorescence (IF)

Culture (most specific diagnostic test) Grown on McKoy cells then stained with iodine

Inoculation of pus into yolk sac of embryonated hens egg and examination of yolk sac membrane for organism.

Inoculation of tissue culture (MacKoys cells)

MacKoy cells stained with iodine Note! inclusions

See semester 1 Herpes Demonstration