수산물 시료에서 병원성 비브리오균의 신속 검출을 위한 현장적용형 전처리 방법 개발

Abstract

Aquatic products provide a favorable environment for microbial proliferation, posing hygiene management issues, and eating raw aquatic products infected with pathogenic Vibrio spp. such as V. parahaemolyticus, V. vulnificus, and V. cholerae causes food poisoning. Currently, molecular biological approaches based on gene amplification are mostly utilized to identify microbiological hazards. The gene amplification approach is extremely reliable, however it has a lengthy processing time and a high cost. This work devised a pretreatment approach for detecting pathogenic Vibrio spp. in aquatic products utilizing PCR with a brief enrichment culture and a simple procedure that included desalting and DNA extraction. The purity and yield of DNA extracted using the new pretreatment approach and DNA extracted using a commercial Kit were compared to establish compatibility. Since then, it has been used to sterile and non-sterile homogenized mussels artificially contaminated with three pathogenic Vibrio spp. at concentrations of 100, 101, and 102 CFU/g to determine optimal DNA extraction conditions. The effectiveness of the pretreatment technique was then tested using 15 different types of aquatic products obtained from Busan, Tongyeong, and Pohang. The results of detecting pathogenic Vibrio spp. by MPN-PCR and the pretreatment technique were compared, and the same findings were found to validate the efficiency of the pretreatment method. This method is intended to be utilized not only for aquatic products, but also for agricultural and livestock products, and it may be used as basic data for the development of molecular biological fast detection based on gene amplification.