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

作者：Greig David R; Hickey Tadgh J; Boxall Megan D; Begum Hafsah; Gentle Amy; Jenkins Claire; Chattaway Marie A

来源：Diagnostic Microbiology and Infectious Disease, 2018, 90(3): 171-176.

DOI：10.1016/j.diagmicrobio.2017.11.017

• Real-time PCR
• Vibrio cholerae
• Serogroup
• Biotype
• Toxin encoding genes

We report the development and validation of a duo-triplex real-time polymerase chain reaction (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 cholera toxin (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), nonfunctional 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.

• 出版日期2018-3