Sidewise-flow test strip detection kit for detecting geese origin component in food and feed and application of kit

Abstract

The invention discloses a nucleic acid rapid detection kit for a geese origin component in food and feed and an application method of the kit, belonging to the fields of molecular biology and immunology. According to the invention, a high-sensitivity high-specificity method of PCR in nucleic acid detection is combined with an immuno gold staining rapid detection technology in immunological detection, extracted target DNA is subjected to specific amplification through designing a specific primer and labeling the primer and the amplified product is combined with a gold labeled antibody immobilized on the test strip in a developing liquid so as to form stable and visible detection zone and quality control zone, thereby realizing the rapid and accurate detection of geese origin component in food 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 goose-derived components in food and feed, and a preparation and application method 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 ruminants 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 Hygiene 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 prod...

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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 "Containing 3' modified oligonucleotides of pseudo-isocytosine nucleic acid base derivatives and their application as primers or probes" provides a method using specially modified nucleotides as substrates 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 semi-quantitative method of adding internal control nucleic acid to pathogenic nucleic acid "Detection method" is to use internal control probes to reduce interference. Among the above methods, the use of hot start technology and circular primers is a common method, and the rest of the methods either use specially synthesized substrates and substrates, or need to introduce 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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