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

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 (bovine spongiformencephalopathy, 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 sources to ruminant feed In 2000, the European Union extended the 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 produc...

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Problems solved by technology

In terms of primer optimization, the invention patent with the publication number CN102146432A - "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. Self-circularization at room temperature to avoid the formation of heterodimers. The invention patent with the publication number CN102719547A - "Reagent for Real-time Fluorescent Quantitative PCR for Detecting HER2 Gene Expression Level" also uses a similar method for real-time quantitative PCR amplification; The invention patent with the publication number CN101842494A - "Using chimeric primers to reduce heterodimer formation" describes a method for amplification using chimeric primers; in the optimization of the reaction substrate, the invention patent with the publication number CN101171343A "3'-Modified Oligonucleotides Containing Pseudo-Isocytosine Nucleobase Derivatives and Their Use as Primers or Probes" provides a method for using specially modified nucleotides as substrates to reduce primer-dimer formation. The method of using probes or introducing internal control probes can also reduce the interference of non-specific amplification. The invention patent with the publication number CN101957373A-"A method for semi-quantitative detection of pathogen nucleic acid by adding internal control nucleic acid" uses internal control probes To reduce interference, in the above methods, the use of hot start technology and circularized primers is the current method, and the rest of the methods either use specially synthesized substrates and substrates, or need to introduce the second hybridization process through probes, which will increase the detection The complexity of the process, especially the probe hybridization method, loses the convenience of the test strip method due to the need for an incubation process

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