A lateral flow test strip detection kit for the detection of deer source components in food and feed and its application

Abstract

The invention discloses a preparation method and an application method of a nucleic acid rapid detection kit for detecting Cervusaxis in rood and feed and belongs to the fields of molecular biology and immunology. According to the invention, a high-sensitivity and high-specificity polymerase chain reaction method in nucleic acid detection and an immunogold staining quick detection technology in immunological detection are combined, a unique primer is designed and marked, specific amplification is carried out on extracted target DNA, and an amplification product is combined with an immunogold labelled antibody on a test strip in a developing solution, thus forming a stable and visible detection strip and quality control strip for quickly and accurately detecting Cervusaxis in rood and feed.

Description

technical field [0001] The invention belongs to the fields of molecular biology and immunology, and relates to PCR nucleic acid amplification of deer source DNA in food and feed and preparation and application methods of a lateral flow immune colloidal gold test strip kit. Background technique [0002] In order to prevent the spread of bovine spongiform encephalopathy (BSE) through beef and beef bones added to animal feed, the European Union and the United States in 1994 and 1997 respectively enacted a ban on adding ruminant to ruminant feed Regulations for protein of animal origin, the European Union in 2000 extended this ban to prohibit feeding processed mammalian, bird and fish protein ingredients to animals intended for food. The "Administrative Measures for the Safety and Sanitation of Animal-derived Feed Products" promulgated by the Ministry of Agriculture of my country in 2004 stipulates that the use of animal-derived feed products (except milk and dairy products) in ...

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Problems solved by technology

In terms of primer optimization, the invention patent with the publication number CN 102146432A - "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 primers self-cyclize at room temperature to avoid the formation of heterodimers. The invention patent with the publication number CN 102719547A - "Reagent for Real-time Fluorescent Quantitative PCR for Detecting HER2 Gene Expression Level" also uses a similar method for the amplification of real-time quantitative PCR. Zeng; the invention patent with the publication number CN 101842494A - "Using Chimeric Primers to Reduce Heterodimer Formation" describes a method of using chimeric primers for amplification; in the optimization of the reaction substrate, the publication number CN The invention patent of 101171343A "3'modified oligonucleotides containing pseudo-isocytosine nucleobase derivatives and their application as primers or probes" provides a method of using specially modified nucleotides as substrates to reduce the number of primers The method of dimer formation, the use of probes or the introduction of internal control probes can also reduce the interference of non-specific amplification. The invention patent with the publication number CN 101957373A "A method for semi-quantitative detection of pathogen nucleic acid by adding internal control nucleic acid" is Internal control probes are used to reduce interference. Among the above methods, hot start technology and circularized primers are common methods. The remaining methods either use specially synthesized substrates and substrates, or need to introduce a second hybridization process through probes. , 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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