Rapid Identification of Vibrio parahaemolyticus in Seafood by Nested PCR
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Abstract:
Vibrio parahaemolyticus is a food-borne pathogen that causes gastroenteritis and other diseases worldwide, with consumption of contaminated raw or undercooked seafood. To develop a method for rapid and specific detection of V. parahaemolyticus in seafood, its genomic sequence was analyzed and compared with those of other Vibrio species, a V. parahaemolyticus-specific genetic marker was screened, and a nested-PCR-based method to rapid detect V. parahaemolyticus was established based on this gene. Additionally, the specificity, sensitivity, and reproducibility of this method were assessed. The results showed that the target amplicon only appeared with V. parahaemolyticus genomic DNA as the template, but not the other 11 Vibrio species and non-Vibrio species. The limits of detection of this method were 10 fg V. parahaemolyticus genomic DNA and 6.6 colony-forming units (CFU) for purified culture. Artificial contamination experiments indicated that V. parahaemolyticus could be detected after two hours of enrichment with an initial concentration of 25.7 CFU/100 mL. In conclusion, the VP1331 gene can be used as a V. parahaemolyticus-specific marker, and this PCR method can be used for the detection and identification of V. parahaemolyticus in contaminated seafood.
