Multiplex fluorescent quantitative PCR method for detecting and identifying three food-borne pathogenic bacteria

Abstract

The invention discloses a multiplex fluorescent quantitative PCR method for detecting and identifying three food-borne pathogenic bacteria, and belongs to the technical field of microbial detection. The invention discloses multiplex fluorescent quantitative PCR amplification primers and probes for detecting and identifying three food-borne pathogenic bacteria, wherein the primers and probes comprise the following sequences: a primer pair and a probe for amplifying an escherichia coli O157:H7rfbE gene as shown in SEQ ID NO:1-3; a primer pair and a probe for amplifying a salmonella invA gene as shown in SEQ ID NO:4-6; and a primer pair and a probe for amplifying a listeria monocytogenes hlyA gene as shown in SEQ ID NO: 7-9; and the sequences as shown in SEQ ID NO:1-9 are used together in one detection. The multiplex fluorescent quantitative PCR method disclosed by the invention can provide methods and data support for accurate and rapid detection of three food-borne pathogenic bacteria including escherichia coli O157: H7, salmonella and listeria monocytogenes in food.

Description

technical field [0001] The invention relates to the technical field of microbial detection, in particular to a multiplex fluorescence quantitative PCR method for detecting and identifying three kinds of food-borne pathogenic bacteria. Background technique [0002] The most commonly used detection method for food-borne pathogens is bacterial isolation and culture combined with biochemical identification. Traditional bacterial identification requires three steps: proliferation culture, selective culture and biochemical identification, which is the "gold standard" for the detection of food-borne pathogens "The disadvantage is that it takes a long time, usually 5-6 days, and the detection sensitivity is low, and quantitative analysis cannot be performed. With the development and application of molecular biology, real-time fluorescent quantitative polymerase chain reaction (qPCR) has become a powerful research and diagnostic tool. Simple, rapid, sensitive and specific are the fou...

AI Technical Summary

Problems solved by technology

[0002] The most commonly used detection method for food-borne pathogens is bacterial isolation and culture combined with biochemical identification. Traditional bacterial identification requires three steps: proliferation culture, selective culture and biochemical identification, which is the "gold standard" for the detection of food-borne pathogens "The disadvantage is that it takes a long time, usually takes 5-6 days, and the detection sensitivity is low, and quantitative analysis cannot be performed

With the development and application of molecular biology, real-time fluorescent quantitative polymerase chain reaction (qPCR) has become a powerful research and diagnostic tool. Simple, rapid, sensitive and specific are the four key aspects of qPCR to meet the needs of nucleic acid detection. , and has been applied to the detection of foodborne pathogens, but the effect on the detection and identification of pathogens is uneven

With the continuous emergence of new food-borne pathogens in recent years, there are currently no reports on simultaneous multiplex fluorescent quantitative PCR detection methods for Escherichia coli O157:H7, Salmonella and Listeria monocytogenes. , there is an urgent need to establish a multiplex fluorescent PCR method that can simultaneously detect three food-borne pathogens of Escherichia coli O157:H7, Salmonella and Listeria monocytogenes, and use it for food testing

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