Rapid detection of Vibrio parahaemolyticus by PCR targeted to the histone-like nucleoid structure (H-NS) gene and its genetic characterization.

ABSTRACT

AIMS: The aim of this study was to explore a new PCR target gene for Vibrio parahaemolyticus, based on the histone-like nucleoid structure (H-NS) gene. METHODS AND RESULTS: Primers for the H-NS gene were designed for specificity to V. parahaemolyticus and incorporated into a PCR assay. The PCR assay was able to specifically detect all of the 82 V. parahaemolyticus strains tested, but did not result in amplification in the 47 other Vibrio spp. and nonVibrio spp. strains. The detection limit of the PCR assay was 0.14 pg purified genomic DNA and 1.8 × 10(5) CFU g(-1) spiked oyster samples from V. parahaemolyticus RIMD2210633. Furthermore, a multiplex PCR assay targeting the hns, tdh and trh genes was successfully developed to detect virulent V. parahaemolyticus strains. CONCLUSIONS: The H-NS-based PCR assay developed in this study was sensitive and specific, with great potential for field detection of V. parahaemolyticus in seawater or seafood samples. SIGNIFICANCE AND IMPACT OF THE STUDY The H-NS gene was validated as a new specific marker gene in PCR assays for accurate detection and identification of V. parahaemolyticus, which has the potential to be applied in diagnostics and taxonomic studies.