Detection of Vibrio cholerae from Aquatic Animals by Dulplex PCR Based on lolB and toxR Genes

V.cholerae is well recognized as pathogenic bacteria that cause diseases in both human and aquatic animals.In this study,two pairs of specific primers were designed based on the lolB and toxR genes,and a dulplex polymerase chain reaction(PCR) was established to detect all V.cholerae serogroup and biotypes.The results indicated that both PCR primer pairs could simultaneously amplify 519 bp and 779 bp gene fragments from the chromosomal DNA of V.cholerae.No positive reaction was detected in 4 control strains of pathogenic Vibrio.The sensitivity of the dulplex PCR assay also showed that the designed primer pairs could detect V.cholerae at a level of 3.42 × 103 CFU/mL.The assay was simple,rapid,specific,sensitive and applicable for the detection of V.cholerae for different purposes.