Preparation method of nucleic acid lateral flow test strip for detecting yersinia enterocolitica and application thereof

Abstract

The invention discloses a preparation method of a quick nucleic acid test strip detection kit for food-borne pathogenic microorganism-yersinia enterocolitica and an application thereof, and also discloses a design method of a primer for specific nucleic acid PCR (polymerase chain reaction) amplification for yersinia enterocolitica, and a method for detecting a PCR amplification product with the nucleic acid test strip for nucleic acid lateral flow and an application. The method disclosed by the invention can be used for detecting the amplification product of the target nucleotide sequence of pathogenic microorganisms from other sources by use of the specific nucleotide sequence amplification product, wherein the specificity is guaranteed by the amplification primer.

Description

technical field [0001] The invention belongs to the fields of molecular biology and immunology, and relates to a preparation and application method of a lateral flow immune colloidal gold test strip kit of a nucleic acid amplification product of Yersinia enterocolitica in food and processing raw materials. Background technique [0002] In order to effectively control foodborne diseases caused by pathogenic microorganisms in food production and import and export trade, and ensure public safety in the food field, it is necessary to develop sensitive, convenient and accurate detection methods for foodborne pathogenic microorganisms. Among the food-borne disease risk factors, microbial food poisoning ranks first among the prevalent food-borne diseases in my country, and among the food-borne pathogenic microorganisms, the most typical pathogenic bacteria is enterocolitis Yersinia Bacteria, Shigella flexneri, Staphylococcus aureus, Listeria monocytogenes, Escherichia coli O157: H7 ...

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Problems solved by technology

In terms of primer optimization, the invention patent with the publication number CN 102146432 A - "A method for reducing the dimer of a pair of partially homogeneous primers" describes a primer design method with a short palindromic sequence at the 5' end of the primer, The primer self-cyclizes at room temperature to avoid the formation of heterodimers. The invention patent with the publication number CN 102719547 A - "Reagent for Real-time Fluorescent Quantitative PCR for Detecting HER2 Gene Expression Level" also uses a similar method for real-time quantitative PCR Amplification; Publication No. CN 101842494 A invention patent - "use chimeric primers to reduce heterodimer formation" describes a method of using chimeric primers to amplify; in the optimization of the reaction substrate, The invention patent of publication number CN 101171343A "3' modified oligonucleotides containing pseudoisocellular nucleobase derivatives and their application as primers or probes" provides a method using specially modified nucleotides as In order to reduce the formation of primer dimers, the use of probes or the introduction of internal control probes can also reduce the interference of non-specific amplification. The publication number is the invention patent of CN 101957373 A-"A method for adding internal control nucleic acid to pathogenic nucleic acid. "Semi-quantitative detection method" uses internal control probes to reduce interference. Among the above methods, the use of hot start technology and circular primers is a common method. The introduction of the needle into the second hybridization process will increase the complexity of the detection process, especially the probe hybridization method, which loses the convenience of the test strip method due to the need for an incubation process

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