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Primer group and kit for detecting origin of meat in food with multiple PCR (Polymerase Chain Reaction)

A primer set and food technology, applied in the field of biological genetic engineering, can solve the problems of long time, poor meat source stability, protein variability, etc., and achieve the effect of improving the efficiency of detection and shortening the time of detection

Inactive Publication Date: 2011-02-02
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the easy denaturation of protein in the process of meat product processing, the stability of ELISA method in identifying the source of meat is poor, prone to deviation, and the detection steps are complicated and time-consuming, which is not conducive to popularization

Method used

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  • Primer group and kit for detecting origin of meat in food with multiple PCR (Polymerase Chain Reaction)
  • Primer group and kit for detecting origin of meat in food with multiple PCR (Polymerase Chain Reaction)
  • Primer group and kit for detecting origin of meat in food with multiple PCR (Polymerase Chain Reaction)

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Experimental program
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Effect test

Embodiment 1

[0016] The invention is for the detection of the meat source in the food, and the product lengths amplified by the PCR reaction are pig: 136bp, cow: 705bp, sheep: 199bp, chicken: 327bp.

[0017] 1. Extraction of DNA from meat products

[0018] Sample processing: Tissue samples of porcine, cattle, sheep and chicken of equal quality were ground and fully ground for later use.

[0019] Extraction of total DNA: take 30mg sample and suspend in 200μl TE, add 400μl lysate (5mol / L CuSCN; 0.05mol / L Tris-HCl, pH6.4; 0.02mol / L EDTA, pH8.0; 1.3% TritonX-100 ), incubate at 70°C for 10 minutes, add an equal amount of chloroform to mix, centrifuge (10000r / min, 5min), take the upper aqueous phase, add an equal amount of ice ethanol to precipitate at -20°C for 10 minutes, centrifuge (10000r / min, 5min), Discard the supernatant, wash once with 70% ethanol, dry in the air, and dissolve the precipitate with 50 μl TE; store at -20°C until use.

[0020] 2. Screening and verification of primers

...

Embodiment 2

[0049] A kit for detecting meat origin in food by multiplex PCR, the kit includes a primer set, a PCR tube and 2×TaqPCR reaction mixed reagents, the primer set consists of a pair of primers for detecting pork, a pair of primers for detecting chicken, and a pair of primers for detecting beef pair and detection of mutton primer pair, the upstream and downstream primers of the pork primer pair are respectively the nucleotide sequences shown in SEQ ID NO.1 and SEQ ID NO.2 in the sequence table, and the upper and lower primers of the detection chicken primer pair The downstream primers are respectively the nucleotide sequences shown in SEQ ID NO.3 and SEQ ID NO.4 in the sequence listing, and the upstream and downstream primers of the beef primer pair are respectively SEQ ID NO.5 and SEQ ID NO.6 in the sequence listing. As shown in the nucleotide sequence, the upstream and downstream primers of the detection mutton primer pair are respectively the nucleotide sequences shown in SEQ ID...

Embodiment 3

[0051] Application of a kit for multiplex PCR detection of meat sources in food:

[0052] Sample DNA extraction, PCR and electrophoresis methods were the same as in Example 1.

[0053] Apply multiplex PCR to the detection of cooked meat products in the market, purchase various cooked meat products from the market, and record the meat quality types identified by their labels, extract total DNA, perform multiplex PCR reactions, and detect the real components.

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Abstract

The invention discloses a primer group and a kit for detecting the origin of meat in food with multiple PCR (Polymerase Chain Reaction). The primer group for detecting the origin of meat in food with multiple PCR comprises a detection pork primer pair, a detection chicken primer pair, a detection beef primer pair and a detection mutton primer pair. The primer group can be applied to a multiple PCR technology for detecting the origin of meat in food. The kit can agilely and quickly determine the origin of meat in cooked meat products, and the multiple PCR method shortens the detection time, improves the detection efficiency, and is applied to the field of food detection.

Description

technical field [0001] The invention belongs to the technical field of biogenetic engineering, and relates to a primer set and a detection kit for multiplex PCR detection of meat sources in food. Background technique [0002] In the current meat market, some units and individuals substitute chicken for pork and pork for beef in order to pursue their own interests, and use low-priced and low-quality meat to pretend to be high-priced and high-quality meat, which greatly damages the interests of consumers and consumers. healthy. Since the country has not stipulated a unified meat testing method, and has not formed a mature, fast and easy industry method for testing the source of meat, the current quality supervision department mainly uses the taste, color and texture of meat products to determine the source of meat. These traditional identification methods have played a very important role in the identification of counterfeit raw meat in the market. However, for many cooked m...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
Inventor 乔建军孙艳华张智禹关丛笑
Owner TIANJIN UNIV
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