Vaginal Wet Mount Interpretation

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D. Microscopic Examination

1. Rapid examination performed to detect motile Trichomonas

2. Specimen swab is placed into a tube containing approximately 1.0ml of saline (0.9% NaCl)

3. Specimen is agitated or twirled for seconds to release the secretions.

4. Another slide is prepared with 10% KOH preparation added to the single drop of sample.

5. A gram stain may also be prepared.

Here copy and paste of the full pdf instructions [1]

Analysis of Vaginal Secretions

I.Introduction: The most common complaint encountered by the health professional working in women’s health is the vaginal discharge or vaginal disorder. The three major causes for these symptoms are bacterial infections, candidiasis, and trichomoniasis. The causative agent for each of these conditions is different; the clinical presentation can be very similar. The treatment for each of these is unique so the diagnosis is critical. In addition, sexual partners are often treated to avoid reinfection.

A. The Clinical Laboratory Improvement Ammendment 1. The classification of direct examination (wet mount) and KOH examination for vaginal secretions as provider-performed microscopy tests were included.

2. When non-laboratory personnel (NP, PA, and MD) perform these tests the designated laboratory director is responsible for assuring the accuracy and reliability of the testing performed. 3. These tests enable health care providers to diagnose and treat the common causes of vaginitis.

B. Specimen Collection and Handling

1.Optimal recovery from a pelvic examination of microorganisms and other elements for microscopic elucidation.

2. Pelvic sample using a non-lubricated speculum, moistened only with warm water.

 a.Lubricants contain anti-microbial contaminates.
 b.Specimen collection device is a polyester (Dacron) tip.

1.Cotton is toxic to many organisms such as Neisseria. 2.Wooden shafts are toxic to Chlaymdia.

c.Wire loop can also be used.

3. Vaginal secretions are collected from the posterior vaginal fornix and the vaginal pool.

4. Sample transported with patient medical history such as menstrual status, exposure to sexually transmitted diseases, and use of lubricants, creams or douches.

5. Tests should be performed immediately; if unavoidably delayed then specimen must be held at room temperature.

a. N. gonorrhoeae and Trichomonas are adversely affected by refrigeration.

b. Refrigeration of Chlaymidia and Herpes is preferred because overgrowth of normal flora is prevented.

C. pH

1.2. The pH of vaginal secretion specimens should be determined using commercial pH paper before placing the swab into saline. Differential diagnosis using the pH of vaginal secretions

a. pH 3.8-4.5 range for healthy vagina; this pH is maintained by lactobacilli in the normal flora that they produce hydrogen peroxide which enhances the acid environment.

b. pH > 4.5 is associated with bacterial vaginosis, trichomoniasis, and atrophic vaginitis.

c. pH < 4.5 limited diagnostic value but excludes Neisseria gonorrhoeae, Gardnerella vaginalis, Haemopholis influenzae, and Trichomonas vaginalis.

D. Microscopic Examination

1.Rapid examination performed to detect motile Trichomonas.

2.Specimen swab is placed into a tube containing approximately 1.0ml of saline (0.9% NaCl).

3.Specimen is agitated or twirled for seconds to release the secretions.

4.Another slide is prepared with 10% KOH preparation added to the single drop of sample.

5.A gram stain may also be prepared.

E. Wet Mount Examination – Direct Examination

1.The Dacron swab sample is put into a 1m tube of saline.

2.Roll the swab onto the sides of the test tube.

3.Remove the swab and a drop is placed on a glass slide.

4.A glass cover slip is placed on the drop of saline-suspended specimen.

5.Examine the sample using brightfield at low-power (10x) used to assess overall distribution and evaluate epithelial cells; amount, type, and groupings.

6.Examine using high-power (40x) magnification to quantify the elements present such as yeast and bacterial morphology.

7.Reportable elements are red and white cells, bacteria, yeast and hyphae/pseudohyphae, trichomonads, clue cells, parabasal and basal cells, and squamous epithelial cells.

F. KOH Preparation and Amine Test

1.Prepare a KOH slide and the amine test by adding 1 drop of 10% KOH directly onto the drop of the specimen on the microscope slide.

2.Check the slide for a “fishy” odor; the name of the test is often called the “whiff” test.

a. The distinct foul-smelling odor is trimethylamine caused by the volitilization product of polyamines, which results from3.

4. 5. 6. 7. II. the alkaline pH change when KOH is added.

b.When microbial flora of the vagina is altered significantly by proliferation of microbes, there is an increase in polyamine production due to the development of the discharge and increased foliation of epithelial cells. Report the Amine test as either positive or negative. KOH preparation occurs at the same time as the direct examination but this slide is set aside to allow the KOH to dissolve the epithelial and blood cells in the specimen. Heat the KOH preparation to speed up the process of digestion. This enhances visualization of any fungal elements present. Low-power (10x) magnification is used to screen the preparation and high-power (40x) magnification is used identify and enumerate fungal elements or pseudohyphae. Limited to fungal identification and Amine testing. Cellular Components

A.Blood Cells 1.White Blood Cells

a.few seen in healthy women

b.increase seen during ovulation and menses

2. Red Blood Cells

a.usually contamination due to menstruation B. Bacterial Flora

1.Lactobacilli – 50% - 90% of bacteria in healthy women

a.large, nonmotile, gram positive rods

b.produce lactic acid as waste product

c.produce hydrogen peroxide

Qualitative semen analysis is approved for the PPM category. b. Quantitative semen analysis includes evaluating the color, appearance, time of liquefaction, volume, motility, count, pH, fructose, viability and the presence of abnormal forms. This analysis is classified as high complexity.