Novel Multiplex Polymerase Chain Reaction Assay to Detect Virulence-related Genes in Vibrio parahaemolyticus
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Abstract:
Two multiplex polymerase chain reaction (PCR) assay systems were developed to detect 11 common virulence-related Vibrio parahaemolyticus (V. parahaemolyticus) genes (toxR, collagenase gene, toxS, trh, tdh, tlh, UreR, FlaA, ompW, AspA, and fur). The two systems were applied to investigate the distribution of these virulence genes in 248 strains of V. parahaemolyticus isolated from the environment and aquatic products. The results showed that FlaA, encoding flagellin protein; ompW, encoding the outer membrane protein; and fur, encoding the ferric uptake regulator were the most widely distributed (100%) genes, followed by the alkaline serine protease gene, AspA (99.60%). Genes with > 90% distribution included collagenase gene; tlh, encoding the thermolabile hemolysin; and the virulence regulatory genes, toxR and toxS, and tlh and toxR possessed similar distribution. The urease encoding UreR was distributed at a very low level (1.21%), while the thermostable direct hemolysin gene, tdh and the thermostable related hemolysin gene, trh were not detected in any of the 248 V. parahaemolyticus strains. The multiplex PCR assays established in this study can be kconsidered as a rapid and efficient method to investigate the distribution of virulence genes, which can provide data for virulence research and ris assessment regarding V. parahaemolyticus.
