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Primer pair and probe for detecting chicken-derived components as well as kit and application of kit

A kit and primer pair technology, applied in biochemical equipment and methods, microbiological determination/inspection, DNA/RNA fragments, etc., can solve the problems of lengthy and limited food and meat detection applications, and achieve accurate results and easy operation Fast, highly sensitive results

Pending Publication Date: 2021-04-27
HANSHAN NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, existing technologies require considerable operational skills, expensive equipment, and lengthy processes, which limit the application of food and meat detection.

Method used

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  • Primer pair and probe for detecting chicken-derived components as well as kit and application of kit
  • Primer pair and probe for detecting chicken-derived components as well as kit and application of kit
  • Primer pair and probe for detecting chicken-derived components as well as kit and application of kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0066] 1. Sample collection

[0067] Use sterile scissors to save the sample to be tested into a sterile centrifuge tube for later use.

[0068] 2. DNA extraction

[0069] The DNA of the meat samples to be tested was extracted with 5% Chelex-100.

[0070] 3. RAA-LFD

[0071] 3.1. Design of primers and probes for specific sequence amplification of chicken-derived components

[0072] Download the complete chicken (NC_040902.1) mitochondrial genome sequence, use the chicken ND4L gene sequence as the target gene, analyze the characteristics of the chicken sequence and compare it with the sequences of common adulterated meat species, including duck (NC_009684.1), cattle ( NC_006853.1), sheep (NC_001941.1), and pig (NC_000845.1), species-specific amplification primers were designed using Primer Premier 5.0 software. The probe sequence is located in the middle section of the amplification primer and modified by tetrahydrofuran. Primer design follows the following principles:

...

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Abstract

The invention discloses a primer pair and probe for detecting chicken-derived components, and discloses a kit designed according to the primer pair and probe. The kit comprises an RAA reaction system and an LFD test strip, wherein the RAA reaction system comprises general dry powder for an RAA reaction, a Tris-HCl buffer solution, an upstream primer JI-F, a downstream primer JI-R, a probe JI-PRO, a sample DNA extracting solution, MgAcO and ddH2O; a quality control line of the LFD test strip is provided with an avidin-colloidal gold specific antibody; and a detection line of the LFD test strip is provided with a biotin antibody and a fluorophore antibody. Based on a recombinase-mediated isothermal nucleic acid amplification technology and a lateral flow immunoassay technology, the invention provides a novel chicken-derived component adulteration on-site detection method; visual detection of naked eye results can be realized; and compared with other detection methods, the method is high in detection sensitivity, simple and rapid to operate, free of special equipment and accurate in result, can meet the detection requirement of field samples, and is especially suitable for field detection of farmer markets, supermarkets, wholesale markets and the like.

Description

technical field [0001] The invention relates to a primer pair and a probe for detecting chicken-derived components and its kit and application, belonging to the field of rapid detection of biotechnology and food safety, in particular to a recombinase-mediated isothermal nucleic acid amplification technology-lateral flow Primer pairs and probes, kits and applications for detection of chicken-derived components by immunotechniques. Background technique [0002] Meat and meat products are considered an important source of protein for the human body and have evolved into an essential dietary ingredient, widely consumed around the world for their appreciated taste and flavor. Unscrupulous producers may try to substitute chicken for beef, lamb, and other expensive meats due to the lure of economic profit (economically motivated adulteration, EMA). Meat adulteration has occurred around the world. The 2013 horsemeat scandal (the so-called "Horsegate" scandal) in the UK and Europe ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12Q1/6844C12N15/11
CPCC12Q1/6888C12Q1/6844C12Q2521/507C12Q2563/107C12Q2563/131C12Q2565/125Y02A50/30
Inventor 郑玉忠林敏林丽云杨培奎黄慧莹孔美兰王忠合
Owner HANSHAN NORMAL UNIV
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