title: Quantification of normal vaginal constituents using a new wet preparation technique.
abstract: OBJECTIVE
  This study aimed to evaluate a new method for preparing vaginal wet preparations to enable quantification of cells and lactobacilli. The current nonstandardized technique allows for a variable amount of vaginal fluid collected, diluted by a variable amount of saline/KOH, and no quantification of constituents.
  
  
  METHODS
  The vaginal fluids from 100 randomly selected women without vulvovaginitis symptoms presenting to the author's practice at Mayo Clinic underwent analysis by the quantification technique. Women were excluded if they were younger than 18 years, had antibiotics within the past 2 months, currently on their period, had placed anything in the vagina for the past 24 hours, used Depo-Provera, or were lactating.
  
  
  METHODS
  All the wet preparations were made by mixing the natural vaginal fluids with 3 mL of sterile normal saline. Spinal diluting fluid was added to the saline preparation. The saline and KOH mixtures were injected into separate wells of KOVA Glasstic Grid Slide and analyzed with a phase-contrast microscope at 40× and 60×. The concentration of leukocytes, lactobacilli, and squamous cells and the degree of maturation of the majority (>50%) of squamous cells were assessed, and it was determined whether there was excessive non-lactobacilli bacteria (EB) as evident by clumps of bacteria in the background fluid and speckling on the squamous cells.
  
  
  RESULTS
  The 3 most common patterns to occur were as follows: First, 51% (95% confidence interval [CI] = 41%-60%) of the total specimens had abundant lactobacilli, no leukocytes, more than 50% fully maturated squamous cells, and no EB. Second, 22% (95% CI = 14%-32%) of the total specimens had low lactobacilli counts, no leukocytes, more than 50% undermaturated squamous cells, and no EB. Third, 12% (95% CI = 6%-20%) of the total specimens had abundant lactobacilli, leukocytes, more than 50% fully maturated squamous cells, and no EB.
  
  
  CONCLUSIONS
  It is imperative to be able to objectively quantify normal vaginal secretion constituents so that (1) the abnormal patterns can be demarcated and (2) treatment targets of what constitutes healthy vaginal conditions can be provided.
authors.affiliation: Mayo Clinic, Scottsdale, AZ 85259, USA. fowler.stuart@mayo.edu
authors.forename: R Stuart
authors.initials: RS
authors.lastname: Fowler
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001419:Bacteria; D058491:Bacterial Load; D001431:Bacteriological Techniques / Q000379:methods*; D001826:Body Fluids / Q000166:cytology* / Q000382:microbiology*; D002452:Cell Count; D003584:Cytological Techniques / Q000379:methods*; D004847:Epithelial Cells / Q000166:cytology; D007887:Fabaceae; D005260:Female; D006801:Humans; D056546:Lactobacillales / Q000145:classification / Q000302:isolation & purification; D007962:Leukocytes / Q000166:cytology; D008875:Middle Aged; D013048:Specimen Handling / Q000379:methods*; D014621:Vagina / Q000166:cytology / Q000382:microbiology / Q000502:physiology*; D055815:Young Adult