Loop-mediated isothermal amplification detection primer groups of Escherichia coli 0157, detection method and reagent kit

Abstract

The invention discloses loop-mediated isothermal amplification detection primer groups of Escherichia coli 0157, a detection method and a reagent kit. A set of specific detection primer groups, the detection reagent kit containing detection primer groups, the method using the detection reagent kit for detection through loop-mediated isothermal amplification and the detection method for determining whether the Escherichia coli 0157 exists in a sample to be tested are designed and screened for rfbE genes of the Escherichia coli 0157. The detection reagent kit and the detection method have the advantages that the sensitivity is high, the specificity is high, the accuracy rate reaches 99% and the detection time is short, only 8-10 hours are spent from sample treatment to result reporting, no polymerase chain reaction instrument or electrophoresis apparatus is required, the operating process is simple, the problems that traditional detection methods are long in consumed time, traditional cultural methods are low in sensitivity, cross reaction exists during serum agglutination and the like are solved, the detection reagent kit and the detection method are significant to timely and effective handling of food-borne sudden public health events, and the detection reagent kit and the detection method are particularly applicable to self-checking of basic level detection mechanisms and food processing enterprises.

Description

Technical field: [0001] The invention belongs to the field of biotechnology, and in particular relates to a detection primer set, a detection method and a kit for a loop-mediated isothermal amplification detection of Escherichia coli O157. Background technique: [0002] Escherichia coli O157 (Escherichia coli O157) is a type of diarrhea-causing Escherichia coli that can cause severe food poisoning, and Escherichia coli O157:H7 is the main serotype of hemorrhagic Escherichia coli. Since 1982, E.coli O157:H7 has successively caused outbreaks and epidemics in the United States, Britain, Canada, Japan and other countries. The pathogen is an important pathogen that causes diarrhea and hemorrhagic enteritis, and 2% to 7% of patients will develop hemorrhagic nephrotic syndrome, and its infection has become a worldwide problem. In addition to E.coli O157:H7, Escherichia coli O157:NM (nonmobile), which produces Shiga toxin and can ferment sorbitol, has become an important pathogen i...

AI Technical Summary

Problems solved by technology

[0004] In recent years, molecular detection methods for E. coli O157 such as PCR and LAMP methods have been gradually established. These methods have high specificity and sensitivity, but due to the complexity of food types, the interference of finished samples and background microorganisms on E. coli O157 is relatively large. In addition, E.coli O157 is greatly affected by factors such as culture temperature, salinity, pH value and various components of the medium. Therefore, there is no complete, efficient and sensitive method among the current methods. The detection method is applicable to samples from all sources including (food, water, soil and clinical samples)

In addition, recent studies have found that the improved EC broth has a certain impact on the recovery and growth of the target bacteria E.coli O157 due to the high content of antibacterial agents such as bile salts and novobiocin, resulting in some complex matrix samples. .coli O157 cannot be checked out

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