Open AccessSeptaplex PCR assay for rapid identification of Vibrio cholerae including detection of virulence and int SXT genes
Author(s) -
Mantri Chinmay K.,
Mohapatra Saswat S.,
Ramamurthy Thandavarayan,
Ghosh Raikamal,
Colwell Rita R.,
Singh Durg V.
Publication year - 2006
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2006.00491.x
Subject(s) - vibrio cholerae , biology , virulence , microbiology and biotechnology , cholera toxin , gene , pilus , ribosomal rna , polymerase chain reaction , intergenic region , genetics , bacteria , genome
Abstract In this study, we describe a septaplex PCR assay for rapid identification of Vibrio cholerae including detection of the virulence and int sxt genes. Conditions were optimized to amplify fragments of ISRrRNA (encoding for 16S–23S rRNA gene, Intergenic spacer regions), O1rfb (O1 serogroup specific rfb), O139rfb (O139 serogroup specific rfb), ctxA (cholera toxin subunit A), tcpA (toxin coregulated pilus), and int sxt (sxt integron) simultaneously in a single PCR. The septaplex PCR was evaluated using 211 strains of V. cholerae and six water samples for in situ testing. PCR results were correlated with genotype data obtained by individual PCR and slot‐blot assays. The one‐step PCR described here can be used to identify V. cholerae accurately and rapidly. Also, the virulence and int sxt genes can be simultaneously detected, providing a useful method for monitoring pathogenic, int sxt‐positive and nonpathogenic, int sxt‐negative V. cholerae serogroups both in the environment and clinical settings.