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The distribution of genes for an outer membrane protein (OmpW) and a regulatory protein (ToxR) inVibrio cholerae and other organisms was studied using respective primers and probes. PCR amplification results showed that all (100%) of the 254V. cholerae strains tested were positive forompW and 229 (∼98%) of 233 were positive fortoxR . None of the 40 strains belonging to otherVibrio species produced amplicons with eitherompW - ortoxR -specific primers, while 80 bacterial strains from other genera tested were also found to be negative by the assay. These studies were extended with representative number of strains usingompW - andtoxR -specific probes in DNA dot blot assay. While theV. cholerae strains reacted withompW probe, only one (V. mimicus ) out of 60 other bacterial strains tested showed weak recognition. In contrast, several strains belonging to otherVibrio species (e.g.,V. mimicus ,V. splendidus ,V. alginolyticus ,V. fluvialis ,V. proteolyticus ,V. aestuarianus ,V. salmonicida ,V. furnissii , andV. parahaemolyticus ) showed weak to strong reactivity to thetoxR probe. Restriction fragment length polymorphism analysis and nucleotide sequence data revealed that theompW sequence is highly conserved amongV. cholerae strains belonging to different biotypes and/or serogroups. All of these results suggest that theompW gene can be targeted for the species-specific identification ofV. cholerae strains. The scope of this study was further extended through the development of a one-step multiplex PCR assay for the simultaneous amplification ofompW andctxA genes which should be of considerable value in the screening of both toxigenic and nontoxigenicV. cholerae strains of clinical as well as environmental origin.

Rapid Method for Species-Specific Identification of Vibrio cholerae Using Primers Targeted to the Gene of Outer Membrane Protein OmpW