Quick preculture method for determining low concentration Vibrio spp. in contaminated seafood and its verification

This study aimed to present a new approach for detecting pathogenic Vibrio spp., including V. parahaemolyticus, V. vulnificus, and V. cholerae in contaminated seafood. This method combined polymerase chain reaction (PCR) with a concise preculturing step and a simplified DNA extraction procedure. The effectiveness of the method was confirmed using contaminated mussel samples. To validate this proposed method, 15 different shellfish samples were analyzed and the results were compared with the results obtained using a conventional method, mostprobable-number-PCR (MPN-PCR). These results confirm the efficiency of this new approach in detecting pathogenic Vibrio spp. This method takes approximately 6 h for results and offers significant time saving with a reduction of 75% compared with MPN-PCR. Additionally, it demonstrated high sensitivity for the efficient Vibrio spp. detection, and the simplification of the DNA extraction process using boiling makes it accessible to nonexperts and contributes to overall time savings in the sample preparation stage. The proposed method in this study suggests a time-efficient and simplified alternative approach for detecting Vibrio spp. in seafood products, providing an in-situ application for improving safety measures in the food industry.