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Procedures of specimen collection(refer to lab

skill On STI), laboratory examinations to
confirm the diagnosis of STI, non-STI vaginal
Neisseria gonorrhoeae
Specimen collection
● Use sterile swabs or loops, prepare smear at
bedside or clinic,and inoculate directly onto culture
media or put immediately in a non-nutritive
transport media, such as Stuart or Amies media
Gram stain
● Microscopy—detects leucocytes and
intracellular Gram negative diplococci
Culture (direct or inoculated from swabs in
transport media)
● Incubation in 5% carbon dioxide atmosphere—
use selective media (such as modified Thayer
Martin media) for 24-48 hours at 35C. Presumptive
colonies are picked for identification
Presumptive identification
● Typical Gram negative diplococci seen on Gram
stain, positive oxidase test
_ lactamase testing
● Chromagenic cephalosporin—rub growth from
five colonies onto rehydrated nitrocefin disc or
emulsify in nitrocefin reagent on slide—a positive
reaction (red colour) detects penicillin resistance
Antimicrobial susceptibility testing
● Minimum inhibition concentration testing or
disc diffusion tests detect trends in resistance to
antibiotics and plasmid carriage

Chlamydia trachomatis
Specimen collection
● Preferable to use cervical or urethral specimens
collected by swab or loop; however, first void urine
then self administered vaginal swabs or tampons can
be used with some detection techniques.
● Use appropriate swabs, non-toxic for tissue
culture and Dacron for nucleic acid detection
methods. Cytobrush samples also give good yields.
● Put swabs immediately in appropriate transport
media. Store cold before transport to the laboratory
Direct microscopy
● Giemsa staining, useful for eye infections but
lacks sensitivity for genital specimens
● Tissue culture with McCoy cells; this used to be
the most sensitive method but is time consuming
and has now been superseded by nucleic acid
detection in many laboratories
Direct immunofluorescence
● Direct antibody staining with fluorescence
labelled antibody, has 92% sensitivity, 97%
specificity in symptomatic men, 79%
sensitivity, and 98% specificity in women with
intermediate prevalence
Enzyme immunoassays
● ELISA techniques to detect Chlamydia
lipopolysaccaride antigens. Sensitivity low: 50-80%
depending upon population and test used.
Specificity is good at around 95%
Nucleic acid detection
● LCR and PCR kits are available. Good
sensitivity and specificity: sensitivity ranges from 95%
in male urine to 98% in female cervical swabs, and
specificity around 99% for all samples
Vaginal discharge infections
The most common causes of vaginal discharge are
Trichomonas vaginalis, Candida, and bacterial
vaginosis. Trichomonas and Candida can be
detected with wet preparation microscopy at the
clinic or by culture methods in a laboratory.

Bacterial vaginosis can be determined with clinical
criteria, Amsel’s score, or by using a Gram staining
method, such as Nugent’s score.
A positive Amsel’s score is based on the presence of
three of four of the following
● Characteristic homogeneous, white-grey,
adherent discharge
● Vaginal pH 4.5
Specimen collection
● Vaginal swab, preferably from the wall of
posterior fornix. Either do direct microscopy
immediately or place swab in transport media, this
must be read within two hours
Wet preparation
● Microscopic examination for the presence of T
vaginalis, Candida, and the presence and
approximate percentage of clue cells
Potassium hydroxide preparation
● Smell for typical amine smell of whiff test,
microscopically examine for Candida
●pH test with pH paper
Gram stain
● Nugent’s score for bacterial vaginosis
T vaginalis or candida culture
● InPouch or Diamond media for T vaginalis,
Sabouraud agar plates for Candida. Feinberg-
Whittington media can be used for both Candida
and T vaginalis