VAGINAL FLUID

Vaginal Secretions
-examined in the clinical laboratory to diagnose infections and complications of pregnancy, as well as
for forensic testing in sexual assault patients.

Vaginitis
-One of the most common conditions diagnosed for female patients, particularly women of childbearing
age.
-characterized by abnormal vaginal discharge or odor, pruritus, vaginal irritation, dysuria, and
dyspareunia.
-Most often, vaginitis is secondary to bacterial vaginosis (BV), trichomoniasis, or vulvovaginal candidiasis
-Can also occur with noninfectious conditions, such as vaginal atrophy, allergies, and chemical irritation

Specimen Collection and Handling

• Collected during pelvic examination
• Speculum moistened with warm water is used to visualize the vaginal fornices
• Lubricants may contain antibacterial agents and must not be used
• Collected by swabbing the vaginal walls and vaginal pool to collect epithelial cells along with the vaginal
secretions
• Sterile polyester-tipped swabs on a plastic shaft are used
• Cotton swabs = toxic to Neisseria gonorrhoeae
• Wood in a wooden shaft = toxic to Chlamydia trachomatis
• Calcium alginate = can inactivate herpes simplex virus (HSV)
• The swab is placed in a tube containing 0.5-1.0 mL of sterile physiological saline. The swab should be
twirled in the saline vigorously to dislodge particulates from the swab. Failure to dislodge particles may
lead to erroneous results
• Alternative: dilute a sample of vaginal discharge in one to two drops of normal saline solution directly
on a microscope slide. Then a second sample is placed in 10% KOH solution in the same manner. Cover
slips are placed over both slides for microscopic examination
• Specimens should be analyzed immediately.
• If delayed in transport or analysis, specimens must be kept at room temperature to preserve the motility
of Trichomonas vaginalis and the recovery of Neisseria Gonorrhoeae
• Refrigerated = C. trachomatis and HSV (to prevent overgrowth of normal flora)
• T. vaginalis = must be examined within 2 hours of collection
Color and Appearance

Normal Findings in Vaginal Secretions

Appearance White, flocculent discharge

pH 3.8-4.2
Amine (whiff) test Negative
WBCs Rare to 2+
Lactobacilli Predominant
Clue Cells Absent
Other Cells Absent (except RBCs during
menses)
Other organisms Other lactobacilli subgroups,
occasional yeast

Clinical Features and Laboratory Findings in Vaginitis

Microscopic Procedures
-Saline wet mounts and KOH mounts = initial screening tests
-Gram stain = confirmatory examination for yeast or Bacterial vaginosis
-
Wet Mount examination
- Typical constituents found in vaginal fluid wet mounts include squamous epithelial cells, WBCs, RBCs,
clue cells, parabasal cells, basal cells, bacteria, motile T. vaginalis, yeast, and hyphae/ pseudohyphae.
- Intravaginal medications might leave oil droplets that can interfere with the interpretation of wet
mounts. In this case, a Gram stain is useful to detect yeast or BV.

Cells found in microscopic examination:

• Squamous Epithelial Cells

• Clue Cells
• White Blood Cells
• Red Blood Cells = can be present during menstruation or due to desquamative inflammatory
process. KOH are used to detect the present of yeast cells and lyse the RBCs
• Parabasal Cells = Round to oval-shaped. 16 to 40 µm in diameter. Ratio of nucleus to cytoplasm is
1:1 to 1:2, with marked basophilic granulation or amorphic basophilic structures (“blue blobs”) in
the surrounding cytoplasm.
They are located in the luminal squamous epithelium of the vaginal mucosa. Increased numbers of
parabasal cells, if present with large numbers of WBCs, can indicate DIV
• Basal Cells = Cells are round. 10 to 16 µm in diameter. Ratio of nucleus to cytoplasm is 1:2. They
are not normally seen in vaginal fluid and, if present and accompanied by large numbers of WBCs
and altered vaginal flora, can suggest DIV
• Bacteria = Lactobacillus spp.-normally comprise the largest portion of vaginal bacteria. Appears as
large, gram-positive, nonmotile rods and produce lactic acid, which maintains the vaginal pH at 3.8
to 4.5. Hydrogen peroxide, produced by lactobacilli subgroups, also can help to suppress the
overgrowth of other organisms. Absent or decreased numbers of lactobacilli relative to the number
of squamous epithelial cells suggests an alteration in the normal flora. Often the lactobacilli are
replaced by increased numbers of any of the following:
o Mobiluncus spp. (thin, curved, gram-negative, motile rods)
o Prevotella spp.
o Porphyromonas spp.
o Bacteroides spp. (anaerobic gram-negative rods)
o Gardnerella vaginalis (short, gram-variable coccobacilli)
 o Peptostreptococcus spp. (gram-positive cocci)
o Enterococcus spp. (gram-negative cocci)
o Mycoplasma hominis
o Ureaplasma urealyticum
• Trichomonas vaginalis
• Yeast Cells

KOH Preparation
1. Prepare a clean glass slide labeled with the patient’s name and unique identifier.
2. Place one drop of vaginal specimen on the slide
3. Add one drop of 10% KOH to the slide
4. Allow the KOH slide preparation to rest for up to 5 minutes to allow cellular tissue and other
debris to dissolve. Gentle heating may speed the dissolving process.
5. Cover the specimen with a cover slip, removing any air bubbles.
6. Examine the slide under the 10× objective for overall assessment and for yeast pseudohyphae.
7. Switch to the 40× objective to examine for budding yeast cells (smaller blastopore blastospore).
8. Record the results.
Amine (Whiff) Test
1. Apply one drop of the saline vaginal fluid suspension to the surface of a clean glass slide
2. Add one drop of 10% KOH directly to the vaginal sample
3. Holding the slide in one hand, gently fan above the surface of the slide with the other hand and
assess for the presence of a fishy amine odor
4. Report as positive or negative
Positive: The presence of a fishy odor after adding KOH
Negative: The absence of a fishy odor after adding KOH

Other diagnostic tests:

• Gram Stain
• Culture
• DNA Testing
• Point of Care Tests
VAGINAL DISORDERS
1. Bacterial Vaginosis
2. Trichomoniasis
3. Candidiasis
4. Desquamative Inflammatory Vaginitis
5. Atrophic Vaginitis
Additional Procedures for Vaginal Secretion Associated with Pregnancy

• Fetal Fibronectin Test

• AmniSure Test
• Actim PROM
• ROM Plus

Reference: Urinalysis and Body Fluids. 7th Edition. F.A. Davis Company. By Susan King Strasinger and Marjorie
Schaub Di Lorenzo.