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. 1996;4(1):2–6. doi: 10.1155/S1064744996000026

Assessment of Vaginal Lactobacillary Flora in Wet Mount and Fresh or Delayed Gram's Stain

Gilbert G G Donders 1,✉, Annie Vereecken 2, Geert Salembier 2, Ben Van Bulck 3, Bernard Spitz 1
PMCID: PMC2364465  PMID: 18476056

Abstract

Objective: The assessment of the vaginal lactobacillary flora helps to direct further diagnostic microbiologic investigations in genital infectious disease and seems to represent a powerful tool in predicting infectious morbidity and preterm labor during pregnancy. In the absence of a “gold standard,” we studied the variations in assessing lactobacillary morphotypes according to the method used.

Methods: The lactobacillary flora from 183 pregnant women was classified according to 3 groups: normal, intermediate, and abnormal. This grading of lactobacilli was appled to vaginal and cervical specimens by means of 1) immediate wet-smear microscopy, 2) Gram's stain on a fresh, air-dried specimen, and 3)delayed Gram's stain after specimen transportation in Stuart's growth medium for 3–6 h.

Results: The assignment of intermediate or abnormal flora (grade II or grade III) showed high concordance rates among the different preparatory techniques, but the assignment of grade I (normal flora) did not. Fewer lactobacilli were found 2.6 times more often after Gram's stains of fresh specimens [Relative Risk (RR) 2.6, 95% confidence interval (CI) 1.7–4.1] and 6 times more often when the Gram%s stain was performed in a delayed examination after transport than in a fresh wet-mount specimen (RR 6.2, 95% CI 2.5–15.6). Disturbed lactobacillary grades were also found more frequently in specimens from the cervix than those from the vagina (RR 4.0, 95% CI, 1.5–10.4).

Conclusions: There are discrepancies in the diagnosis of lactobacillary grades between gram-stained and fresh vaginal specimens. The evidence is ambiguous as to which of the 2 methods is responsible. If an evaluation is to be done on a gram-stained specimen, then the storage of the sample in Stuart transport medium before staining should be avoided.

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Selected References

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  1. DUNKELBERG W. E., Jr DIAGNOSIS OF HEMOPHILUS VAGINALIS VAGINITIS BY GRAM-STAINED SMEARS. Am J Obstet Gynecol. 1965 Apr 1;91:998–1000. doi: 10.1016/0002-9378(65)90569-7. [DOI] [PubMed] [Google Scholar]
  2. Domingue P. A., Sadhu K., Costerton J. W., Bartlett K., Chow A. W. The human vagina: normal flora considered as an in situ tissue-associated, adherent biofilm. Genitourin Med. 1991 Jun;67(3):226–231. doi: 10.1136/sti.67.3.226. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Donders G. G., Moerman P., De Wet G. H., Hooft P., Goubau P. The association between Chlamydia cervicitis, chorioamnionitis and neonatal complications. Arch Gynecol Obstet. 1991;249(2):79–85. doi: 10.1007/BF02390366. [DOI] [PubMed] [Google Scholar]
  4. Donders G., De Wet H. G., Hooft P., Desmyter J. Lactobacilli in Papanicolaou smears, genital infections, and pregnancy. Am J Perinatol. 1993 Sep;10(5):358–361. doi: 10.1055/s-2007-994761. [DOI] [PubMed] [Google Scholar]
  5. Eschenbach D. A., Davick P. R., Williams B. L., Klebanoff S. J., Young-Smith K., Critchlow C. M., Holmes K. K. Prevalence of hydrogen peroxide-producing Lactobacillus species in normal women and women with bacterial vaginosis. J Clin Microbiol. 1989 Feb;27(2):251–256. doi: 10.1128/jcm.27.2.251-256.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. GARDNER H. L., DUKES C. D. Haemophilus vaginalis vaginitis: a newly defined specific infection previously classified non-specific vaginitis. Am J Obstet Gynecol. 1955 May;69(5):962–976. [PubMed] [Google Scholar]
  7. GRAY L. A., BARNES M. L. VAGINITIS IN WOMEN, DIAGNOSIS AND TREATMENT. Am J Obstet Gynecol. 1965 May 1;92:125–136. doi: 10.1016/0002-9378(65)90113-4. [DOI] [PubMed] [Google Scholar]
  8. HUNTER C. A., Jr, LONG K. R. A study of the microbiological flora of the vagina. Am J Obstet Gynecol. 1958 Apr;75(4):865–871. doi: 10.1016/0002-9378(58)90668-9. [DOI] [PubMed] [Google Scholar]
  9. Hay P. E., Lamont R. F., Taylor-Robinson D., Morgan D. J., Ison C., Pearson J. Abnormal bacterial colonisation of the genital tract and subsequent preterm delivery and late miscarriage. BMJ. 1994 Jan 29;308(6924):295–298. doi: 10.1136/bmj.308.6924.295. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hillier S. L., Krohn M. A., Nugent R. P., Gibbs R. S. Characteristics of three vaginal flora patterns assessed by gram stain among pregnant women. Vaginal Infections and Prematurity Study Group. Am J Obstet Gynecol. 1992 Mar;166(3):938–944. doi: 10.1016/0002-9378(92)91368-k. [DOI] [PubMed] [Google Scholar]
  11. Lindner J. G., Plantema F. H., Hoogkamp-Korstanje J. A. Quantitative studies of the vaginal flora of healthy women and of obstetric and gynaecological patients. J Med Microbiol. 1978 Aug;11(3):233–241. doi: 10.1099/00222615-11-3-233. [DOI] [PubMed] [Google Scholar]
  12. Martius J., Krohn M. A., Hillier S. L., Stamm W. E., Holmes K. K., Eschenbach D. A. Relationships of vaginal Lactobacillus species, cervical Chlamydia trachomatis, and bacterial vaginosis to preterm birth. Obstet Gynecol. 1988 Jan;71(1):89–95. [PubMed] [Google Scholar]
  13. Mårdh P. A., Westtöm L. Adherence of bacterial to vaginal epithelial cells. Infect Immun. 1976 Mar;13(3):661–666. doi: 10.1128/iai.13.3.661-666.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Ofek I., Beachey E. H. Mannose binding and epithelial cell adherence of Escherichia coli. Infect Immun. 1978 Oct;22(1):247–254. doi: 10.1128/iai.22.1.247-254.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Påhlson C., Larsson P. G. The ecologically wrong vaginal lactobacilli. Med Hypotheses. 1991 Oct;36(2):126–130. doi: 10.1016/0306-9877(91)90253-u. [DOI] [PubMed] [Google Scholar]
  16. Sobel J. D., Schneider J., Kaye D., Levison M. E. Adherence of bacteria to vaginal epithelial cells at various times in the menstrual cycle. Infect Immun. 1981 Apr;32(1):194–197. doi: 10.1128/iai.32.1.194-197.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Spiegel C. A., Amsel R., Holmes K. K. Diagnosis of bacterial vaginosis by direct gram stain of vaginal fluid. J Clin Microbiol. 1983 Jul;18(1):170–177. doi: 10.1128/jcm.18.1.170-177.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]

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