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Primer pair and probe for detecting sheep-derived component, kit and application thereof

A sheep-derived, kit-based technology, applied in biochemical equipment and methods, microbiological determination/inspection, DNA/RNA fragments, etc., can solve the problems of restricting the application of food and meat detection, lengthy and other problems, and achieve simple and fast operation, Accurate results and high detection sensitivity

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 sheep-derived component, kit and application thereof
  • Primer pair and probe for detecting sheep-derived component, kit and application thereof
  • Primer pair and probe for detecting sheep-derived component, kit and application thereof

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 sheep-derived components

[0072] Download the complete mitochondrial genome sequence of sheep (NC_001941.1), use the sequence of the sheep D-loop gene as the target gene, analyze the characteristics of the sheep sequence and compare it with the sequences of common adulterated meat species, including duck (NC_009684.1), Cattle (NC_006853.1), pig (NC_000845.1), chicken (NC_040902.1), 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 a probe for detecting a sheep-derived component, and discloses a kit designed according to the primer pair and the probe. The kit comprises an RAA reaction system and an LFD test strip; the RAA reaction system comprises RAA reaction general dry powder, Tris-HCl buffer solution, an upstream primer YANG-F, a downstream primer YANG-R, a probe YANG-PRO, sample DNA extracting solution, MgAcO and ddH2O; and an avidin-colloidal gold specific antibody is arranged on a quality control line of the LFD test strip, and a biotin antibody and a fluorophore antibody are arranged on a detection line of the LFD test strip. According to the primer pair and the probe for detecting the sheep-derived component, a novel sheep-derived component adulteration field detection method is provided based on a recombinase-mediated isothermal nucleic acid amplification technology and a lateral flow immune technology, so that visual detection of naked eye results can be realized, and compared with other detection methods, the method is high in detection sensitivity, simple, convenient and rapid to operate, does not need special equipment, and the method is especially suitable for field detection in farmer markets, supermarkets, wholesale markets and the like.

Description

technical field [0001] The invention relates to a primer pair and a probe for detecting sheep-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 the detection of sheep-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 other inexpensive meats for lamb 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 is one of the mos...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12Q1/6844C12N15/11
CPCC12Q1/6888C12Q1/6844C12Q2521/507C12Q2563/107C12Q2563/131C12Q2565/125
Inventor 林敏林丽云郑玉忠杨培奎王忠合孔美兰黄慧莹
Owner HANSHAN NORMAL UNIV
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