SEXUALLY TRANSMITTED

INFECTIONS – LAB
DIAGNOSIS
Pathogen
Bacterial infections
1.Neisseria gonorrhoeae
2. Chlamydia trachomatis
3. Chlamydia trachomatis (serovars L1–L3)
4. Treponema pallidum
5. Haemophilus ducreyi
6. Klebsiella (Calymmatobacterium)
granulomatis
7. Mycoplasma genitalium
Clinical manifestations and other associated diseases
GONORRHOEA
Men: urethral discharge (urethritis), epididymitis, orchitis, infertility
Women: cervicitis, endometritis, salpingitis, pelvic inflammatory disease, infertility,
preterm rupture of membranes, perihepatitis; commonly asymptomatic
CHLAMYDIAL INFECTION
Men: urethral discharge (urethritis), epididymitis, orchitis, infertility
Women: cervicitis, endometritis, salpingitis, pelvic inflammatory disease, infertility,
preterm rupture of membranes, perihepatitis; commonly asymptomatic Both sexes:
proctitis, pharyngitis, Reiter’s syndrome Neonates: conjunctivitis, pneumonia
LYMPHOGRANULOMA VENEREUM
Both sexes: ulcer, inguinal swelling (bubo), proctitis
SYPHILIS
Both sexes: primary ulcer (chancre) with local adenopathy, skin rashes, condylomata
lata; bone, cardiovascular, and neurological damage
Women: pregnancy wastage (abortion, stillbirth), premature delivery
Neonates: stillbirth, congenital syphilis
CHANCROID
Both sexes: painful genital ulcers; may be accompanied by bubo
DONOVANOSIS (GRANULOMA INGUINALE)
Both sexes: nodular swellings and ulcerative lesions of the inguinal and anogenital areas
Men: urethral discharge (nongonococcal urethritis)
Women: cervicitis, endometritis, probably pelvic inflammatory disease
Men: urethral discharge (nongonococcal urethritis)
Women: cervicitis, endometritis, probably pelvic inflammatory disease
VIRAL
Human immunodeficiency virus (HIV) ACQUIRED IMMUNODEFICIENCY SYNDROME (AIDS)
Both sexes: HIV-related disease, AIDS

Herpes simplex virus type 2 Herpes simplex virus type 1 GENITAL HERPES
(less common) Both sexes: anogenital vesicular lesions and ulcerations
Neonates: neonatal herpes (often fatal)

Human papillomavirus GENITAL WARTS

Men: penile and anal warts; carcinoma of the penis Women:
vulval, anal, and cervical warts, cervical carcinoma, vulval
carcinoma, anal carcinoma Neonates: laryngeal papilloma

Hepatitis B virus VIRAL HEPATITIS

Both sexes: acute hepatitis, liver cirrhosis, liver cancer

Cytomegalovirus CYTOMEGALOVIRUS INFECTION

Both sexes: subclinical or nonspecific fever, diffuse lymph
node swelling, liver disease, etc
Molluscum contagiosum virus MOLLUSCUM CONTAGIOSUM
Both sexes: genital or generalized umbilicated, firm skin
nodules
Kaposi sarcoma associated herpesvirus (human herpesvirus KAPOSI SARCOMA
type 8 Both sexes: aggressive type of cancer in immunosuppressed
persons
Protozoal infections

Trichomonas vaginalis TRICHOMONIASIS

Men: urethral discharge (nongonococcal urethritis); often asymptomatic
Women: vaginosis with profuse, frothy vaginal discharge; preterm birth,
low-birth-weight babies Neonates: low birth weight

Fungal infections

Candida albicans CANDIDIASIS

Men: superficial infection of the glans penis Women: vulvo-vaginitis with
thick curd-like vaginal discharge, vulval itching, or burning

Parasitic infestations

Phthirus pubis PUBIC LICE INFESTATION

Sarcoptes scabiei SCABIES
Syphilis (TREPONEMA PALLIDUM)

• Laboratory diagnosis of syphilis consists of demonstration of

treponemes, detection of antibodies and PCR.
Syphilis
• Dark Ground Microscopy
• Direct IF staining ( DFA- TP)
• Silver impregnation method – Levaditi stain (tissue section) , Fontana
stain (smear)

• Culture : Not cultivable, maintained in rabit testes

Serology : Ab detection

• Non Treponemal /STS (standard test for Syphilis)/ / Lipoidal Tests : detect a characteristic non-specific antibody
(called reagin antibody) in the sera of syphilitic patients by using cardiolipin antigen extracted from beef heart.
VDRL,RPR,TRUST,USR

• Specific / Treponemal test:

Specific antibodies are detected by using T. pallidum antigens
TPI (Treponema pallidum immobilization test)
FTA-ABS (Fluorescent treponemal antibody absorption test)
TPA (T. pallidum agglutination test)
TPHA (T. pallidum hemagglutination test)
TPPA (T. pallidum particle agglutination test)

PCR
Chancroid (Haemophilus ducreyi)
• Direct microscopy: H. ducreyi is a pleomorphic gramnegative
coccobacillus; occurs in groups or in parallel chains

• A multiplex PCR assay has been developed for simultaneous detection

of common agents of STIs including H. ducreyi (targeting 16S rRNA).
Lymphogranuloma venereum (LGV)
• Diagnosis of LGV is usually based on serology; biopsy is
contraindicated as there is a risk of sinus tract formation
• NAAT: Nucleic acid amplification test C. trachomatis will be positive
• Antibody detection by ELISA or microimmunofluorescence (MIF)
• Direct detection of inclusion bodies by direct IF or for culture
confirmation
Herpes genitalis
Laboratory diagnosis:
• Staining of scrapings from the base of the lesions with Giemsa’s (Tzanck preparation), or
Papanicolaou’s stain can detect giant cells or intranuclear inclusions of HSV infection
• Viral antigen (by direct IF) or viral DNA (by PCR) can be detected in scrapings from lesions
• Isolation of the virus in scrapings from lesions.
Donovanosis
• Direct microscopy: Smears can be examined after a rapid Giemsa or
Wright’s stain Donovan bodies can be seen as large cyst like
macrophages filled with deeply stained capsulated bacilli having a
safety-pin (bipolar) appearance
• These cysts eventually rupture releasing the bacilli
• They are non-motile, capsulated and gram-negative bacilli.
• Culture: They can be grown on egg yolk medium and on HEp-2 cell lines
• Molecular method: PCR has been developed to differentiate Klebsiella
granulomatis from other Klebsiella species by detecting unique base
changes in the phoE gene.
Gonococcal urethritis (Neisseria
gonorrhoeae)

• Microscopy Gram staining of urethral exudates reveals gram-negative intracellular kidney-shaped

diplococci

• CultureThayer Martin medium (chocolate agar added with antibiotics)

• Molecular Method Nucleic acid amplification tests (NAATs) such as PCR are available for detection
of N. gonorrhoeae from the clinical specimens targeting 16s or 23s rRNA gene.
Chlamydia trachomatis
• Serovars D–K are associated with— genital tract infections
• Serovars L1–L3 causes a sexually transmitted infection,
lymphogranuloma venereum (LGV). It is an ulcerative genital disease
• Specimen: Depends on the type of lesions
• Microscopy: Detects chlamydial inclusion bodies
• Gram staining, Lugol’s iodine and other stains such as Castaneda,
Machiavello or Gimenez stains
• Direct IF: Used for direct detection of inclusion bodies.
• Antigen detection (LPS antigens): By enzyme immunoassays
• Culture: It was the gold standard method in the past
• Egg (yolk sac), mice inoculation and cell line culture
• Cell lines of choice- McCoy, HeLa (for C. trachomatis), HEp2 (for C.
pneumoniae
• Nucleic acid amplification tests (NAAT), e.g. PCR
• The most sensitive and specific method
• Currently the diagnostic assay of choice.
Serology (antibody detection): CFT or ELISA using group specific LPS
antigen
• Micro-IF test detects antibody against species and serovar specific
MOMP antigen.
Urogenital mycoplasma infections
• Mycoplasma (M. hominis, M. genitalium) and Ureaplasma (U.
urealyticum and U. parvum)

• Culture and PCR

Trichomoniasis
• It is the most common parasitic sexually transmitted infection (STI)
Laboratory Diagnosis
• Direct Microscopy Vaginal, urethral discharge, urine sediment and
prostatic secretions can be examined.
• Wet (saline) mount of fresh samples (within 10–20 minutes of
collection) should be done to demonstrate the jerky motile
trophozoites and pus cells.
• Other staining methods include permanent stains (e.g. Giemsa and
Papanicolaou stain), acridine orange, fluorescent stain.
• Direct fluorescent antibody test (DFA).
• Culture :
• Antigen detection in Vaginal Secretion
• Antibody detection
• Molecular methods
Bacterial Vaginosis
• Nugent’s score: It is a scoring system followed for the diagnosis of
bacterial vaginosis; done by counting the number of G. vaginalis,
Mobiluncus and lactobacilli present in the Gram stained smear of
vaginal discharge. A score of more than or equal to 7 is diagnostic.

• Culture: gram-negative (appears gram-variable in smears), nonmotile,

small pleomorphic rod, which shows metachromatic granules
Vaginal Candidiasis
• Candida albicans is the most common species to cause vaginal
candidiasis , followed by C. glabrata and C. tropicalis.

• Laboratory diagnosis
Culture of vaginal secretions on Sabouraud dextrose agar (pasty or dry
white colonies)
Identification by conventional (e.g. germ tube test) or automated
methods (VITEK or MALDI-TOF)
Thank You