A real-time multiplex PCR for the identification and typing of Vibrio cholerae

Publication date: Available online 1 December 2017
Source:Diagnostic Microbiology and Infectious Disease
Author(s): David R Greig, Tadgh J Hickey, Megan D Boxall, Hafsah Begum, Amy Gentle, Claire Jenkins, Marie A Chattaway
We report the development and validation of a duo-triplex real-time PCR for the rapid identification and typing of Vibrio cholerae. The PCR assay targets a species specific toxR gene present in all strains of V. cholerae and used as a marker for the species, wbeO1 and wbfO139, encoding the O1 and O139 somatic antigens, and ctxA encoding CT. The two tcpA variants associated with the Classical and El-Tor biotypes are used to infer biotype. The assay was evaluated using 178 isolates comprising eight different Vibrio species, including 122 isolates of V. cholerae. The PCR results of 171/178 (96.1%) isolates were concordant with the serotyping, biotyping and expected CT results. Variants of toxR (n=3), non-functional wbeO1 (n=1) and CT negative isolates of V. cholerae O1 (n=3) were likely explanations for the mismatched results. This duo-triplex real-time PCR is a reproducible and robust assay for the rapid identification and typing of V. cholerae belonging to the highly pathogenic, pandemic lineages.



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