Seek consent.

Explain procedure.

Use a sterile needle and syringe. Draw up 5 ml of blood from a vein. Put in a plain test tube.

Let test tube sit 20 minutes to allow serum to separate. (Or centrifuge 3�5 minutes at 2000�3000 rpm). In the separated sample, serum will be on top.

Use sampling pipette to withdraw some of the serum. Take care not to include any red blood cells from the lower part of the separated sample.

Hold the pipette vertically over a test card circle. Squeeze teat to allow one drop (50 �l) of serum to fall onto a circle. Spread the drop to fill the circle using a toothpick or other clean spreader.

Attach dispensing needle to a syringe. Shake antigen.* Draw up enough antigen for the number of tests done (one drop per test).

Holding the syringe vertically, allow exactly one drop of antigen to fall onto each test sample. Do not stir.

Rotate the test card smoothly on the palm of the hand for 8 minutes.** (Or rotate on a mechanical rotator.)

After 8 minutes rotation, inspect the card in good light. Turn or tilt the card to see whether there is clumping (reactive result). Most test cards include negative and positive control circles for comparison.

Example
test card

1. Non-reactive (no clumping or only slight roughness)�Negative for syphilis

2. Reactive (highly visible clumping)�Positive for syphilis

3. Weakly reactive (minimal clumping)�Positive for syphilis

NOTE: Weakly reactive can also be more finely granulated and difficult to see than this illustration

* Make sure antigen was refrigerated (not frozen) and has not expired.

** Room temperature should be 22.8^oC�29.3^oC

• Determine if the woman and her partner have received adequate treatment.
• If not, treat woman and partner for syphilis with benzathine penicillin.
• Treat newborn with benzathine penicillin.
• Follow-up newborn in 2 weeks.
• Counsel on safer sex.

RPR

RPR titre

TPHA

Active infection

+

>1:8

+

Latent syphilis

+

Often <1:4

+

False positive

+

Usually <1:4

-

Successful treatment

+ or -

2 titres decrease
(e.g. from 1:16 to 1:4)

+

BV can be diagnosed using simple clinical criteria with or without the aid of a microscope.

Collect specimen

Note colour and consistency of discharge. Take a sample of discharge from the side walls or deep in the vagina where discharge pools (or use discharge remaining on speculum). Touch pH paper to discharge on swab or speculum and note pH.

Prepare slide

Place specimen on a glass slide. Add a drop of 10% potassium hydroxide and note any odour.

What to look for

The diagnosis of BV is based on the presence of at least 3 of the 4 following characteristics

• Homogeneous white-grey discharge that sticks to the vaginal walls

• Vaginal fluid pH >4.5

• Release of fishy amine odour from the vaginal fluid when mixed with 10% potassium hydroxide (positive whiff test)

• "Clue cells" visible on microscopy

Important

Look for evidence of other vaginal or cervical infections�multiple infections are common.

Direct microscopic examination of vaginal discharge can aid in diagnosis of yeast infection (Candida albicans), bacterial vaginosis and trichomoniasis.

Collect specimen

Take a sample of discharge with a swab from the side walls or deep in the vagina where discharge accumulates.

Prepare slide

Mix specimen with 1 or 2 drops of saline on a glass slide and cover with a coverslip.

What to look for

Examine at 100X magnification and look for typical jerky movement of motile trichomonads.

Examine at 400X magnification to look for yeast cells and trichomonads.

To make identification of yeast cells easier in wet mount slides, mix the vaginal swab in another drop of saline and add a drop of 10% potassium hydroxide to dissolve other cells.

Important

Look for evidence of other vaginal or cervical infections�multiple infections are common.

Collect specimen

A Gram stain slide can be prepared at the same time as the wet mount by rolling the swab on a separate slide.

Prepare slide

1. Heat fix.

2. Stain with crystal violet (60 seconds) and rinse.

3. Stain with iodine (60 seconds) and rinse.

4. Decolorize with acetone-ethanol for few seconds (until the liquid runs clear).

5. Stain with safranin (60 seconds) and rinse.

6. Gently blot dry and examine under oil immersion (1000X).

What to look for

1. Lactobacilli only�normal

2. Mixed flora, mainly lactobacilli with a few short rods (coccobacilli)�considered normal

3. Presence of clue cells; mixed flora, mainly Gardnerella-like and anaerobic bacteria with a few lactobacilli�treat for BV

4. Presence of clue cells; mixed flora of Gram-positive, Gram-negative and Gram-variable rods; no lactobacilli�treat for BV

Important

Look for evidence of other vaginal or cervical infections�multiple infections are common.