Lateral flow test paper strip detection kit for detection of fox source components in food and feed and application thereof

Abstract

The invention discloses a preparation method and an application method of a nucleic acid rapid detection kit for detection of fox (VulpesFox) components in food and feeds, and belongs to the field of molecular biology and immunology. According to the preparation method and the application method, a high sensitivity and high specificity method of polymerase chain reaction for nucleic acid detection and an immune colloidal gold rapid detection technology of immunological detection can be combined, a unique primer is designed, the primer is marked, extracted target DNA is specifically amplified, an amplified product is combined with a gold labeled antibody fixed on a test paper strip in a developing solution for formation of a stable and visible detection band and a quality control band, so that fast and accurate detection of the fox source components in the food and feeds can be realized.

Description

technical field [0001] The invention belongs to the fields of molecular biology and immunology, and relates to PCR nucleic acid amplification of fox-derived DNA 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 products) in ...

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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 number is CN 101842494 A invention patent - "Using Chimeric Primers to Reduce Heterodimer Formation" describes a method of using chimeric primers for amplification; in the optimization of the reaction substrate, The invention patent of publication number CN 101171343A "3' modified oligonucleotides containing pseudoisocytosine 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 invention patent with the publication number of CN 101957373 A-"A semi-quantitative method for 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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