Simultaneous Detection of Vibrio alginolyticus, Vibrio parahaemolyticus, Vibrio vulnificus and Vibrio cholerae Using Multiplex PCR

Objective)The objective of this study is to develop a multiplex PCR assay that can simultaneously detect Vibrio alginolyticus, Vibrio parahaemolyticus, Vibrio vulnificus and Vibrio cholerae in the presence of an internal amplification control (IAC).(Method)Species-specific PCR primers were designed based on gyrB gene for Vibrio alginolyticus, collagenase gene for Vibrio parahaemolyticus, vvhA gene for Vibrio vulnificus and ompW gene for Vibrio cholerae, 16S rRNA gene of bacteria as IAC primers was used to indicate false-negative results. Multiple PCR method was developed after optimization reaction condition. (Result)The multiple PCR method was proved to be rapid, high-throughput, sensitive and specific and the existence of IAC could successfully eliminate false-negative results. The multiplex PCR was validated with 69 suspicious Vibrio strains and the results were consistent with the physiological and biochemical experiments.(Conclusion) The multiple PCR method is specific, stable and reliable for the detection of Vibrio alginolyticus, Vibrio parahaemolyticus, Vibrio vulnificus and Vibrio cholerae.