A highly sensitive and specific multiplex PCR assay for simultaneous detection of Vibrio cholerae, Vibrio parahaemolyticus and Vibrio vulnificus.

NEOGI, S.B.; CHOWDHURY, N.; HINENOYA, A; HALDAR, S.; SAIDI, S. M.; KOGURE, K.; LARA, R.J.; YAMASAKI, S.

LETTERS IN APPLIED MICROBIOLOGY

WILEY-BLACKWELL PUBLISHING, INC

Lugar: Londres; Año: 2010 vol. 51 p. 293 - 300

0266-8254

Aim: To develop an effective multiplex PCR for simultaneous and rapid detectionof Vibrio cholerae, Vibrio vulnificus and Vibrio parahaemolyticus, the threemost important Vibrio species that can cause devastating health hazards amonghuman.Methods and Results: Species-specific PCR primers were designed based ontoxR gene for V. cholerae and V. parahaemolyticus, and vvhA gene for V. vulnificus.The multiplex PCR was validated with 488 Vibrio strains including 322V. cholerae, 12 V. vulnificus, and 82 V. parahaemolyticus, 20 other Vibriospecies and 17 other bacterial species associated with human diseases. It coulddetect the three target bacteria without any ambiguity even among closelyrelated species. It showed good efficiency in detection of co-existing targetspecies in the same sample. The detection limit of all the target species was tencells per PCR tube.Conclusions: Specificity and sensitivity of the multiplex PCR is 100% each andsufficient for simultaneous detection of these potentially pathogenic Vibriospecies in clinical and environmental samples.Significance and Impact of the Study: This simple, rapid and cost-effectivemethod can be applicable in a prediction system to prevent disease outbreakby these Vibrio species and can be considered as an effective tool for both epidemiologistand ecologist.