Direct wet mount

1
Direct wet mount KOH preparations
2

• Microscopic observation of unfixed
• wet mounts of clinical specimens,
• either stained or unstained can be
• Useful for the rapid detection of
• Bacterial,fungal and parasitic
• Organisms
• (based on morphology motility)

3
Specimen type

• 1-Swabs of the vaginal mucosa
• vaginal pool
• 2- Scraping of suspected skin area

4
Specimen handling

• Swabs should be placed into tubes containing 0.5
  ml sterile physiologic saline
• ( no refrigeration )
• -Note
• Specimens from the vaginal tract should be tested
  with PH paper before being placed in saline

5
Testing procedure

• - Prepare KOH saline slides
• - Check the KOH slide immediately for a
• fishy amine odor
• - Examine the saline,KOH slides for epithelial
• cells any budding yeast or psuedohyphae
• and quantitate them

6
Micro. Components in KOH prep.

• -Squamous cells
• - WBC
• - RBC
• - Bacteria parasites
• - Yeasts fungi
• - Clue cells

7
Quantitation scheme

• Rare lt 10 org. or cells /
  slide
• Occasional lt 1 org. or cell /10 hpf
• 1(very scanty ) lt1 org. or cell / hpf
• 2(scanty ) 1-5 org. or cells/hpf
• 3( moderate ) 6-30 org. or cells / hpf
• 4( profuse ) gt30 org. or cells / hpf

8
Bacterial vaginosis

• - Etiology
• 1 - Polymicrobial infection
• 2 Marked reduction in normally
• predominant lactobacillus

9
Bacterial vaginosis

• Diagnostic Amsel criteria
• A.Vaginal PH gt4.7
• sensitivity
• specificity (false positive with
  menses
• semen, Trichomona and cervical
  mucus) B. Clue cells (on gt20 of cells )
• C. Positive whiff test (Amine test )
• D. Discharge characteristics
• ( thin,non-clumping,gray-white,adheren
  t )

10
Clue cell

• Diagnostic cells which have small gram negative,
  curved gram negative and gram variable rods
  covering not only on the surface of the cell but
  also spread out past the boundaries , obscuring
  the cytoplasmic margins
• -Adding KOH lava flow appearance

11
(No Transcript)
12
Reference values

• WBC 2
• Lactobacilli predominant
• Clue cells absent
• Other cells absent (except RBCs
• during menses )
• Other org. Other lactobacilli subgroups,
  occasional yeast
• PH lt4.5