Side direction current test strip detection kit for detecting canis familiaris component in feed and application thereof

Abstract

The invention discloses a preparation method of a rapid nucleic acid detection kit for detecting canis familiaris components in feed and an application method thereof, and belongs to the fields of molecular biology and immunology. According to the invention, a high sensitivity and high specificity method of polymerase chain reaction in nucleic acid detection and an immuno gold staining rapid detection technology in an immunological detection are combined, a specific primer is designed and labeled, the extracted target DNA is subjected to specific amplification, and amplified product is combined with a gold labeled antibody immobilized on a test paper tape, so as to form a stable and visible detection band and quality control band, thereby achieving the rapid and correct detection of the canis familiaris components in the main feed.

Description

technical field [0001] The invention belongs to the fields of molecular biology and immunology, and relates to the PCR nucleic acid amplification of animal source raw materials from dogs in feed, and the 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 ...

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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 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 kind of adding internal control nucleic acid to semi-transfer pathogenic nucleic acid Quantitative detection method” means 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 require probe The introduction of 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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