Detection process for food-borne pathogenic bacteria

Abstract

The invention discloses a method for detecting food-borne pathogens and relates to a molecular identification for bacteria, in particular to a method utilizing probe codes to detect the food-borne pathogens, which is a real-time PCR method identifying a plurality of the food-borne pathogens through a single reaction. The invention provides a detection method for the food-borne pathogens which canrealize the typing to genes of 15 templates on a four-color real-time PCR instrument. DNA of a cultivated strain sample is extracted; an universal primer is designed and the sequence measurement of aPCR product is performed; a fluorescent double-stranded displacing probe is designed; a multi-color probe coding system is designed; a temperature-changing hybridization reaction is performed; a single real-time PCR system is constructed; a multiple multi-color real-time PCR system is constructed; and the multiple multi-color real-time PCR system is validated.

Description

Methods for the detection of foodborne pathogens technical field The invention relates to a molecular identification of bacteria, in particular to a method for detecting food-borne pathogenic bacteria using probe codes, which is a real-time PCR method for identifying multiple food-borne pathogenic bacteria through a single reaction. Background technique Traditional bacterial identification mainly relies on methods such as bacterial culture, serology, biochemistry and bacterial morphology for classification and identification ([6], GraciasKSandMcKillipJL.Areviewofconventionaldetectionandenumerationmethodsforpathogenicbacteriainfood[J].CanJMicrobiol, 2004, 50(11):883-890). Although the traditional method is reliable, it often takes days or even weeks to obtain the results; and the phenotype of the bacteria will be affected by the environment, which brings difficulties to the identification; in addition, when the bacteria are in the "VBNC" state, the traditional method is dif...

AI Technical Summary

Problems solved by technology

However, due to the narrow melting point range of PCR products, it is difficult for the melting point curve analysis method to accommodate the simultaneous analysis of more than 5 products, and it is easy to be interfered by non-specific amplification such as primer dimers, making it difficult to judge the results

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