Multiplex PCR–capillary electrophoresis–SSCP used to identify foodborne pathogens

Abstract

Multiplex PCR–CE–SSCP approach was used to identify foodborne pathogens. 16srRNA system including three target fragments localized at different position of 16srRNA gene and gyrB system including two target fragments at gyrB gene were established. SSCP profile was analyzed in a similar way as RAPD and the identity of each peak was determined by relative position to inner standard. Twenty-five reference bacteria strains representing 2 classes, 12 genus and 19 species were tested. All bacteria could be distinctly determined at genus level by 16srRNA system. GyrB system improved discriminating resolution of some bacterial at species level. Phylogenetic analysis showed that 16srRNA based phylogenetic relationship was consistent with traditional classification of these organisms. Good reproducibility and resolution make multiplex PCR–CE–SSCP protocol a promising approach for fast screening of foodborne pathogens.