Establishment of multiple-PCR for diagnosing three food-borne pathogenic bacteria in aquatic products

摘要

A rapid multiplex-PCR method was established in order to detect three foodborne pathogenic bacteria including Salmonella typli , Vibrio parahaemolyticus and Listeria monocytogenes in aquatic products. Three pairs of oligonucleotide primers were designed for multiplex-PCR amplification according to gene coding invasion protein A of Salmonella typli, gene coding immune response of Vibrio parahaemolyticus and regulation histone gene of Listeria monocytogenes. The amplified fragment sizes of these three bacteria were 213 bp, 369 bp and 517bp, respectively. The specificity of the multiplex- PCR was high and the minimum detection limit reached 10~3 CFU/mL. The multiplex-PCR method was used to analyze three aquatic product samples compared with national standard methods, the coincidence rate of two methods reached 100%. The method developed in this study had high sensitivity and specificity, which could be applied for the rapid detection and molecular epidemiology survey of food-borne pathogenic bacteria in aquatic products.