Fourteen-food-borne pathogenic bacterium multiplex PCR detection primer set and kit

Abstract

The invention discloses a fourteen-food-borne pathogenic bacterium multiplex PCR detection primer set and a kit. The detection primer set is composed of primer pairs for detecting salmonella, Shigella, Vibrio parahemolyticus, campylobacter jejuni, campylobacter coli, staphylococcus aureus, bacillus cereus, Listeria monocytogenes, yersinia enterocolitica, enterobacter sakazakii, Escherichia coli, vibrio cholerae, Escherichia coli O157, aeromonas hydrophila and internal positive control. A multiplex PCR detection method based on an ordinary PCR platform is built, multiplex PCR reactions are carried out on genomic DNA, extracted from a sample to be tested, of bacteria in the same reaction system through the primer pairs acquired through analysis and design, and whether the food-borne pathogenic bacteria are contained in the sample or not is judged through the electrophoretic analysis of reaction products.

Description

technical field [0001] The invention belongs to the field of microorganism detection, and relates to a multiplex rapid PCR detection primer set and a kit for fourteen kinds of food-borne pathogenic bacteria. Background technique [0002] Foodborne pathogenic microorganisms are an important biological factor causing food poisoning. With the country's emphasis on food safety, rapid screening of food poisoning causes has become a top priority. The design of the kit of the present invention can be used to screen common diarrhea-causing food-borne microorganisms, including Salmonella, Shigella, Vibrio parahaemolyticus, Campylobacter jejuni, Campylobacter coli, Staphylococcus aureus, cereus Bacillus, Listeria monocytogenes, Yersinia enterocolitica, Enterobacter sakazakii, Escherichia coli, Vibrio cholerae, Escherichia coli O157 and Aeromonas hydrophila , a total of 14 kinds of foodborne pathogenic microorganisms, the test samples come from clinical specimens or the enrichment so...

AI Technical Summary

Problems solved by technology

[0006] The multiplex PCR technology based on the liquid chip detection platform and the multifinder multiplex PCR technology based on double-probe hybridization can realize the rapid screening of multiple detection targets, but currently none of these technologies can detect 14 kinds of food-borne pathogens The microbial kit was successfully developed

Moreover, the multiplex PCR technology based on the liquid-phase chip detection platform performs the PCR reaction in two steps in a PCR reaction tube, and the design of low-concentration specific primers in the first round of PCR leads to a significant decrease in the sensitivity of the PCR reaction, thereby affecting the sensitivity of the entire detection reagent; There is a problem of poor stability in the detection of PCR products by multi-color microsphere labeling

Moreover, the purchase cost of the instrument is high, and the reagent consumables are expensive

[0007] The multifinder multiplex PCR technology based on double-probe hybridization uses two adjacent specific probes to hybridize, which improves the detection specificity and affects the sensitivity of the screening, which is prone to false negative results; this technology uses a next-generation sequencing platform to detect PCR products , such as ABI3130, ABI3500, etc., the reagent consumables are expensive; the detection process goes through steps such as nucleic acid extraction, double-probe hybridization, probe connection, PCR amplification, and product analysis. Use of next-generation sequencing platforms

[0008] Although the multiplex PCR technology based on the arm-PCR principle solves the problems of operation automation and product contamination, the technology requires a large investment and expensive consumables

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