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Indian Journal of Medical Microbiology
Medknow Publications on behalf of Indian Association of Medical Microbiology
ISSN: 0255-0857
EISSN: 0255-0857
Vol. 30, No. 1, 2012, pp. 39-43
Bioline Code: mb12008
Full paper language: English
Document type: Research Article
Document available free of charge

Indian Journal of Medical Microbiology, Vol. 30, No. 1, 2012, pp. 39-43

 en A rapid and reliable species-specific identification of clinical and environmental isolates of Vibrio cholerae check for this species in other resources using a three-test procedure and recA polymerase chain reaction
Roozbehani, A D.; Bakhshi, B; Pourshafie, M R. & Katouli, M

Abstract

Purpose: Vibrio cholerae check for this species in other resources , the cause of cholera, is one of the leading causes of morbidity and mortality in many developing countries. Most laboratories initially rely on biochemical tests for a presumptive identification of these strains, followed by a polymerase chain reaction (PCR)-based method to confirm their identification. The aim of this study is to establish a rapid and reliable identification scheme for V. cholerae using a minimal, but highly specific number of biochemical tests and a PCR assay.
Materials and Methods: We developed a species-specific PCR to identify V. cholerae, using a housekeeping gene recA, and used that to evaluate the sensitivity and specificity of 12 biochemical tests commonly used for screening and / or presumptive identification of V. cholerae in the clinical and environmental samples.
Results: Here we introduced a combination of three biochemical tests, namely, sucrose fermentation, oxidase test, and growth in trypton broth containing 0% NaCl, as also the PCR of the recA gene, for rapid identification of V. cholerae isolates, with 100% sensitivity and specificity. The established method accurately identified a collection of 47 V. cholerae strains isolated from the clinical cases (n = 26) and surface waters (n = 21), while none of the 32 control strains belonging to different species were positive in this assay.
Conclusion: The triple-test procedure introduced here is a simple and useful assay which can be adopted in cholera surveillance programs for efficient monitoring of V. cholerae in surface water and fecal samples.

Keywords
Housekeeping gene, identification, polymerase chain reaction, recA, Vibrio cholera

 
© Copyright 2012 Indian Journal of Medical Microbiology.
Alternative site location: http://www.ijmm.org

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