Method for performing high-flux quick detection on food-borne pathogens by using multiplex PCR (polymerase chain reaction) technique

Abstract

The invention discloses a method for performing high-flux quick detection on food-borne pathogens by using a multiplex PCR (polymerase chain reaction) technique. Primers suitable for multiplex PCR are designed according to conserved region sequences of four main food-borne pathogens salmonella, Staphylococcus aureus, Listeria monocytogenes and Escherichia coli O157:H7, and the multiplex PCR technique is utilized to detect the main food-borne pathogens in the agricultural product. The detection method has the advantages of favorable specificity, higher sensitivity and high detection limit (up to 10<3>copies / ml), and is quick and convenient; and after the primers perform non-cross reaction with 8 similar strains, the amplified strip is clear. The primers and multiplex PCR system can be prepared into a kit for quickly detecting food-borne pathogens; and the kit can greatly shorten the time and enhance the detection efficiency.

Description

technical field [0001] The invention belongs to the field of food safety detection, and in particular relates to a high-throughput rapid detection method for food-borne pathogenic bacteria by using multiplex PCR technology. Background technique [0002] In my country, Salmonella, Staphylococcus aureus, Listeria monocytogenes, and Escherichia coli are widely distributed in the environment. However, most of the daily food testing standards in my country only include sensory indicators and physical and chemical indicators, and lack microbial limit indicators. The food hygiene standards for beef only stipulate the detection of E. coli and salmonella, which is far from the requirements of international standards. Big. Only 12% of my country's overall food standards adopt the standards of the International Codex Alimentarius Commission (CAC), and only 40% adopt the standards of the Food Technical Committee of the International Organization for Standardization. Moreover, 80% of my ...

AI Technical Summary

Problems solved by technology

Enzyme-linked oligonucleotide adsorption assay (PCR-ELISA) has been developed as a detection system for large-scale amplification of bacterial nucleic acids, but samples must be carefully processed to avoid contamination of PCR products

[0009] Traditional foodborne pathogenic microorganism qualitative and quantitative PCR detection technology can only detect a single foodborne pathogenic microorganism at a time, but for foodborne pathogenic microorganism components in processed agricultural products with unclear components of foodborne pathogenic microorganisms, It needs multiple detection methods and multiple experiments to determine, which has a long cycle, heavy workload and high cost

In the future detection of food-borne pathogenic microorganisms in processed agricultural products, these two detection techniques are increasingly unable to meet the special needs of dealing with several, dozens, or dozens of food-borne pathogenic microorganisms at one time, especially large quantities. When processed agricultural products enter commercial production, more effective and rapid detection methods are needed

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