[Establishment and application of a multiplex TaqMan qPCR method for rapid detection of diarrheagenic Escherichia coli].

Abstract

To address the cumbersome operation, long time consumption, and false positive results of biochemical and PCR methods in the detection of diarrheagenic Escherichia coli (DEC), we designed primers and probes targeting 11 virulence genes harbored by five pathotypes of DEC. We then established a novel multiplex TaqMan qPCR method for the rapid detection and typing of DEC after optimization of the experimental conditions. The detection results demonstrated that this method could be used for specific identification and typing of DEC among various foodborne pathogenic strains. Furthermore, it can achieve effective detection when the DNA concentration is no less than 2.5 ng/μL or the culture duration is no less than 18 h. These findings indicate that the developed detection method boasts high specificity, high sensitivity, and short time consumption, which make it applicable for the industrial-scale rapid detection of DEC in food. This study provides novel technical support for food safety sector, which is conducive to further reducing labor input and improving detection efficiency.