Detection of pathogenic Vibrio parahaemolyticus isolated from aquatic animals by dulplex PCR
水产动物致病性副溶血弧菌双重PCR 检测方法的研究

V. parahaemolyticus is a significant pathogen for the aquaculture industry, particularly devastating impact on cultivated Litopenaeus vannamei. In this study, two pair of specific primers was designed that allowed amplification of 285 bp and 368bp gene fragments based on gyrB and toxR genes. A simple dulplex polymerase chain reaction (PCR) that will detect V. parahaemolyticus were established. Consequently, two PCR primers could simultaneously amplify 285 bp and 368bp gene fragments from chromosomal DNA of V. parahaemolyticus in one PCR reaction, and no cross reaction was detected in 4 other pathogenic Vibrio species tested. The results of sensitivity of dulplex PCR showed that the two primers could detect V. parahaemolyticus at a level of as few as 8.8672×103CFU/mL, and detect purified chromosomal DNA at a level of as few as 0.0293 mg/L . Dominant V. parahaemolyticus could be isolated from the positive sample of dulplex PCR through detecting mysis, aquatic products and tank water samples. The PCR protocol amplifying gyrB and toxR gene fragments of the V. parahaemolyticus was established and could be useful in the specific and rapid diagnose of the disease caused by V. parahaemolyticus.