Method for detecting horsemeat in food

A technology for horse meat and food, applied in the field of detection of horse meat components in food using PCR technology, can solve problems such as endangering life and health, lack of sound and unified rapid detection standards for cooked meat products, and harming the interests of consumers, etc., to overcome excessive fragments Effect

Active Publication Date: 2015-01-21
神州基因科技(天津)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Some food manufacturers are opportunistic, shoddy, and cut corners for their own self-interest. While harming the interests of consumers, they seriously endanger people's lives and health.
In addition, there is no sound and unified rapid detection sta

Method used

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  • Method for detecting horsemeat in food
  • Method for detecting horsemeat in food
  • Method for detecting horsemeat in food

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] The 6 pairs of primers provided by the present invention, the length of the target fragment amplified by PCR reaction is as follows, No. 1 primer pair: 183bp; No. 2 primer pair: 202bp; No. 3 primer pair: 231bp; No. 4 primer pair: 266bp; No. 5 primer pair: Primer pair: 268bp; primer pair No. 6: 300bp.

[0019] 1. Genomic DNA extraction of meat products

[0020] Process the sample into minced meat, take 30mg and add it to a 1.5ml EP tube; add 500μl lysate (1M tris-Hcl pH8.0; 0.5M EDTA pH8.0; 5M Nacl; 0.5% N-Lanroyl Sarc0sine; proteinase K; RNaseA ) at 70°C for 0.5-1h or overnight at 50°C, centrifuge (12000r / min, 5-15min), take the supernatant; add an equal volume of phenol, chloroform and isoamyl alcohol (25:24:1) for extraction; take Add 40μl Nacl and 1ml ice ethanol to 400μl water phase, place on ice for 30min; centrifuge (12000r / min, 5min), discard supernatant; wash 3 times with 1ml70% absolute ethanol, dry in the air; add 70-100μlTE, let stand for 30min .

[0021] ...

Embodiment 2

[0055] Validation of commercially available processed horse meat products

[0056] 1. Extracting the genomic DNA of horse meat: the method is the same as that of extracting the genomic DNA of meat products in Example 1.

[0057] 2. PCR verification:

[0058] Using the aforementioned genomic DNA as a template, PCR was performed with the 6 pairs of primers provided in the present invention.

[0059] Reaction system: 20 μl (template: 1 μl; dNTP 0.4 μl; 10×PCR buffer 21 μl; upstream and downstream primers 0.8 μl; rTaq 0.4 μl; ddH 2 O14.6 μl).

[0060] Reaction conditions:

[0061] Step 1: 94°C for 5 minutes;

[0062] Step 2: 94°C for 30s, 62°C for 30s, 72°C for 30s, 30 cycles

[0063] The third step: 5min at 72°C.

[0064] 3. Agarose gel electrophoresis verification

[0065] 2% agarose gel was prepared, and the gel preparation process was the same as that described in Example 1. Voltage 50V, electrophoresis 1h, PCR amplification results were analyzed by gel phase formation...

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Abstract

The invention relates to a method for detecting horsemeat in food. The invention provides a primers for detecting 6 pairs of different fragments horsemeat in food, and the nucleotide sequences are shown as SEQ ID NO.1, NO.2, NO.3, NO.4, NO.5, NO.6, NO.7, NO.8, NO.9, NO.10, NO.11 and NO.12. The invention also provides a kit containing the above primer group. The kit of the invention can fast and accurately detect whether a meat product is doped with horsemeat, and has especially obvious effect in detection of depth processing meat. The invention also provides flexible changeable objective fragments for food inspection personnel to contrast and detect other meat sources, thereby improving the efficiency of detection. In addition, the primers provided by the invention can also be used as primers for real-time fluorescence quantitative PCR to conduct quantification on the horsemeat ingredient doped in food, for better application in food field.

Description

technical field [0001] The invention belongs to the field of food testing and the technical field of molecular biology, and in particular relates to a method for detecting horsemeat components in food using PCR technology. Background technique [0002] In mid-January 2013, the United Kingdom and Ireland found that beef burgers sold in some supermarkets were adulterated with horse meat and other meats. Of the 27 burgers tested, 10 contained horse meat. Since then, the incident of "selling horse meat with a cow's head" has continued to expand, and many European countries have been involved in the scandal. Consumers' trust in the food industry has suffered a serious blow. The EU has also launched "DNA testing" on processed beef from member states. "In order to eliminate the "horse meat storm" worries and restore consumer confidence. [0003] In my country, in recent years, food safety issues have been strongly reflected by the masses and paid close attention to by the society....

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6888C12Q2600/166
Inventor 聂凌云魏迪程东庆池振奋高敬张莹莹赵永良聂凌虎
Owner 神州基因科技(天津)有限公司
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