Preparation method and application of nucleic acid lateral flow test strip for detecting Shigella

Abstract

The invention discloses a rapid nucleic acid test strip detection kit for detecting food-borne pathogenic microorganism Shigella flexneri and an application method thereof, and belongs to the fields of molecular biology and immunology. A preparation method of the rapid nucleic acid test strip detection kit comprises the following steps: combining high sensitivity of polymerase chain reaction in a nucleic acid test, a high-specificity method and an immunity colloid gold rapid detection technology in immunological detection, marking a unique designed primer, carrying out specific amplification on an extracted target DNA (deoxyribonucleic acid), and combining an amplification product and a gold labeling antibody fixed on a test strip in a developing solution, thus forming a detection band and a quality control band which are stable and visible. The rapid nucleic acid test strip detection kit can be used for realizing rapid and accurate detection of main food-borne pathogenic microorganisms.

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 Shigella flexneri 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 risk factors of foodborne diseases, microbial food poisoning ranks first among the prevalent foodborne diseases in my country, and among the foodborne pathogenic microorganisms, the most typical pathogenic bacteria are Shigella, Pathogens such as Enterobacter sakigens, Staphylococcus aureus, Listeria monocytogenes, Escherichia coli O157:H7, etc. ...

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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 of 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 patent for invention - "using 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" means using internal control probes to reduce interference. Among the above methods, the use of hot start technology and circularized 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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