Application ofgroELgene for the species-specific detection ofVibrio parahaemolyticusby PCR

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Aims:Vibrio parahaemolyticus is a significant cause of human gastrointestinal disorders and is transmitted through ingestion of raw or undercooked contaminated seafood. We used the groEL gene for the species-specific detection of V. parahaemolyticus from artificially inoculated shellfish, fish and seawater.Methods and Results:The nucleotide sequences of 24 Vibrio and seven non-Vibrio spp. were compared, and less conserved regions were selected for the designing of primer sets. To detect V. parahaemolyticus specifically, PCR conditions were standardized and tested to evaluate the specificity of primers. A 510-bp band was appeared only from V. parahaemolyticus by PCR. Notably, the detection was shown to be functional at high annealing temperature above 68°C. The groEL primers detected 100 pg and 1 ng of DNA purified from V. parahaemolyticus culture and artificially infected oyster tissue, respectively.Conclusions:The groEL gene is a potential marker for the species-specific detection of V. parahaemolyticus and could be used to detect this bacterium in contaminated food by PCR.Significance and Impact of the Study:PCR using primers designed from groEL gene provide an efficient method for the accurate identification of V. parahaemolyticus from contaminated samples.

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