Establishment and Preliminary Application of the Duplex PCR to Detection of Vibrio vulnificus and Vibrio harveyi

As Vibrio vulnificus and Vibrio harveyi mainly affects marine cultured animals,it is necessary to establish a rapid detection method.According to the cytolysin gene sequence of V.vulnificus and toxR gene sequence of V.harveyi,two pairs of primers were designed and synthesized.The diagnosis of duplex polymerase chain reaction(PCR) for rapid identification of V.vulnificus and V.harveyi was established by optimization of PCR conditions and by testing specificity and sensitiveness.The duplex PCR test results showed that two specific and amplified fragments were produced which sizes were 128 bp for V.vulnificus and 211 bp for V.harveyi,and no cross-reaction tests were done with Streptococcus agalactiae,Streptococcus iniae,Bibrio parahemolyticus,and Vibrio damsela.The lowest concentrations of 12 cfu/mL for V.vulnificus and 18 cfu /mL for V.harveyi could be detected.The duplex PCR would be a specific and sensitive method which was timesaving,laborsaving,quick and high-efficient for identification of duplex V.vulnificus and V.harveyi.