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Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1997 Jul;35(7):1856–1858. doi: 10.1128/jcm.35.7.1856-1858.1997

Comparison of direct-plating and enrichment methods for isolation of Vibrio cholerae from diarrhea patients.

M Lesmana 1, E Richie 1, D Subekti 1, C Simanjuntak 1, S E Walz 1
PMCID: PMC229856  PMID: 9196208

Abstract

A direct-plating method on thiosulfate citrate bile salts sucrose agar (DIR-TCBS) in conjunction with enrichment in alkaline peptone water (APW) incubated for both 6 h and 24 h followed by subculture onto TCBS (APW6h-TCBS and APW24h-TCBS, respectively) was performed on 16,034 rectal swab samples for isolating Vibrio cholerae. A total of 2,932 (18.3%) rectal swab samples were positive for V. cholerae O1 biotype El Tor, with the Ogawa serotype constituting 99.2% of the isolates. There were no significant differences in V. cholerae O1 isolation rates between the three culture systems nor between the combinations of any two systems. However, direct plating plus enrichment demonstrated a significantly higher V. cholerae O1 isolation rate than DIR-TCBS alone (P < 0.02). Conversely, enrichment procedure, alone or in combination with DIR-TCBS, yielded significantly more (P < 0.0001) V. cholerae non-O1 isolates than DIR-TCBS alone. The length of incubation time of the enrichment broth, 6 h, offers no significant advantages over 24 h for the isolation of V. cholerae O1 and non-O1. A 24-h enrichment broth incubation period has the practical advantage of being easy to integrate into a normal laboratory workday, whereas 6-h broth enrichment, although more commonly recommended, requires that arrangements be made for after-hours subculture.

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Selected References

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  1. Barrett T. J., Blake P. A., Morris G. K., Puhr N. D., Bradford H. B., Wells J. G. Use of Moore swabs for isolating Vibrio cholerae from sewage. J Clin Microbiol. 1980 Apr;11(4):385–388. doi: 10.1128/jcm.11.4.385-388.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. DeWitt W. E., Gangarosa E. J., Hug I., Zarifi A. Holding media for the transport of Vibrio cholerae from field to laboratory. Am J Trop Med Hyg. 1971 Sep;20(5):685–688. doi: 10.4269/ajtmh.1971.20.685. [DOI] [PubMed] [Google Scholar]
  3. FEELEY J. C. Isolation of cholera vibrios by positive-recognition plating procedures. J Bacteriol. 1962 Oct;84:866–867. doi: 10.1128/jb.84.4.866-867.1962. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Gangarosa E. J., Dewitt W. E., Huq I., Zarifi A. Laboratory methods in cholera: isolation of Vibrio cholerae (el tor and classical) on TCBS medium in minimally equipped laboratories. Trans R Soc Trop Med Hyg. 1968;62(5):693–699. doi: 10.1016/0035-9203(68)90121-1. [DOI] [PubMed] [Google Scholar]
  5. Large epidemic of cholera-like disease in Bangladesh caused by Vibrio cholerae O139 synonym Bengal. Cholera Working Group, International Centre for Diarrhoeal Diseases Research, Bangladesh. Lancet. 1993 Aug 14;342(8868):387–390. [PubMed] [Google Scholar]
  6. Lesmana M., Rockhill R. C., Sutanti D., Sutomo A. An evaluation of alkaline peptone water for enrichment of Vibrio cholerae in feces. Southeast Asian J Trop Med Public Health. 1985 Jun;16(2):265–267. [PubMed] [Google Scholar]
  7. Pazzaglia G., Lesmana M., Tjaniadi P., Subekti D., Kay B. Use of vaginal tampons in sewer surveys for non-O1. Vibrio cholerae. Appl Environ Microbiol. 1993 Aug;59(8):2740–2742. doi: 10.1128/aem.59.8.2740-2742.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Rennels M. B., Levine M. M., Daya V., Angle P., Young C. Selective vs. nonselective media and direct plating vs. enrichment technique in isolation of Vibrio cholerae: recommendations for clinical laboratories. J Infect Dis. 1980 Sep;142(3):328–331. doi: 10.1093/infdis/142.3.328. [DOI] [PubMed] [Google Scholar]
  9. Spira W. M., Ahmed Q. S. Gauze filtration and enrichment procedures for recovery of Vibrio cholerae from contaminated waters. Appl Environ Microbiol. 1981 Oct;42(4):730–733. doi: 10.1128/aem.42.4.730-733.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]

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