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Detecting method for safely and quickly identifying porcine-derived materials in food and kit for detecting method

A detection kit and detection method technology are applied in the field of detection methods and kits for the safe and rapid identification of porcine-derived components in food, which can solve problems such as the inability to meet the needs of meat product species identification, and achieve reliable technical support, The effect of increased sensitivity, speed and accuracy

Inactive Publication Date: 2015-11-25
乌鲁木齐欧易生物医学科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These illegal acts of passing off pork as beef or mutton, passing fakes as real, or adulterating fakes, cannot meet the needs of meat species identification only by morphological examination of meat tissue

Method used

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  • Detecting method for safely and quickly identifying porcine-derived materials in food and kit for detecting method
  • Detecting method for safely and quickly identifying porcine-derived materials in food and kit for detecting method
  • Detecting method for safely and quickly identifying porcine-derived materials in food and kit for detecting method

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] A detection method for safely and quickly identifying pig-derived components in food, specifically comprising the following steps:

[0049] (1) Design primers: Design pig-specific primers that can only amplify pig mitochondrial DNA. The specific primers include upstream primers and downstream primers, and their sequences are as follows:

[0050] Upstream primer F: 5-ATGAAACATTGGAGTAGTCCTACTATTTACC-3;

[0051] Downstream primer R: 5-CTACGAGGTCTGTTCCGATATAAGG-3;

[0052] (2) Template DNA extraction: The template DNA was extracted by the phenol-chloroform method, and detected by agarose electrophoresis with a mass fraction of 0.8%, and the extracted template DNA was stored at -20°C;

[0053] (3) PCR amplification primers and reaction system: The total reaction system is 25uL, including 12.5uL of PCRMix, 2.5pmol / L each of the upstream primer and downstream primer prepared in step (1), 1μL each, and the template DNA extracted in step (2) 50~ 100ng, water supplement;

[00...

Embodiment 2

[0058] A detection kit for safe and rapid identification of pig-derived components in food, including specific primers, DNA extraction reagents, PCRMix and water. After the four substances are mixed, the total volume of the PCR reaction system mixture is 25uL. Wherein, the specific primers include upstream primers and downstream primers, 1 uL each of the upstream primers and downstream primers; the DNA extraction reagents include five parts of A solution, B solution, C solution, D solution, and E solution. 1ml of solution A; the amount of PCRMix used is 12.5ul; water makes up the rest.

[0059] The PCRMix in the PCR reaction system mixed solution is 0.5 times of the total mixed solution volume; the upstream primer and the downstream primer are each 1 / 25 of the total mixed solution volume; the total amount of DNA is 50-100ng; if the volume of the above components is insufficient The total volume, the insufficient part can be made up with water. Among them, PCRMix is ​​the PCR ...

Embodiment 3

[0073] The method of using the detection kit of the above-mentioned embodiment 2 is as follows:

[0074] (1) Use of DNA extraction reagents:

[0075] (1) Cut the sample to be tested into pieces, add 1ml of A solution per gram of sample, vortex for 30 seconds, place at room temperature for 5 minutes, and centrifuge at 12000rpm for 3 minutes, take the supernatant into a new centrifuge tube;

[0076] (2) Add the supernatant (500ul-800ul) obtained in step (1) into 500ul of B solution, mix it upside down, centrifuge at 12000rpm for 3 minutes, and take the supernatant into a new centrifuge tube;

[0077] (3) Add an equal volume of C solution to the supernatant obtained in step (2) (the amount used is the same volume as the supernatant obtained in step (2), for example: the supernatant obtained after centrifugation in step (2) is 300ul, then add C liquid is 300ul), invert and mix well, centrifuge at 12000rpm for 3 minutes, take the supernatant into a new centrifuge tube;

[0078] (...

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Abstract

The invention discloses a detecting method for safely and quickly identifying porcine-derived materials in food and a kit for the detecting method. The detecting method comprises the following steps of porcine specific primer designing, extraction of template DNA (deoxyribose nucleic acid), preparation of PCR (polymerase chain reaction) amplification primers and a reaction system, and PCR amplification and amplification product detection. The kit comprises a specific primer probe, a DNA extracting reagent, PCR reaction system mixed liquor, agarose powder and the like. The detecting method has the advantages that results are accurate, sensitivity is high, pork ingredients in the food can be detected quickly, illegal behaviors of counterfeit beef and mutton, sham and adulteration are completely eradicated, edible safety of Moslem food of areas inhabited by ethnic minorities such as the Xinjiang Uygur Autonomous Region and the Ningxia Hui Autonomous Region in China and peripheral Central Asia is guaranteed, and a reliable technical support is provided for social stability of China.

Description

technical field [0001] The invention relates to a food detection method, in particular to a detection method for safely and quickly identifying pig-derived components in food and a kit thereof, belonging to the field of food detection methods. Background technique [0002] With the development of the economy and the improvement of people's living standards, the market demand for beef and mutton will continue to expand. In recent years, in order to obtain high profits, some criminals have appeared on the market with fake beef mixed with pork. Fake meat rolls and sliced ​​meat products such as mutton, mutton, etc., and some "fake broths" made of pork head meat, lean pork meat, star anise, cinnamon, pepper and other seasonings have also appeared. These illegal acts of passing off pork as beef or mutton, passing fakes as real, and adulterating fakes, can no longer meet the needs of identifying the species of meat products only by morphological examination of meat tissue. In ord...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 张超刘军陈慕芝焦谊闻娅雷蕊
Owner 乌鲁木齐欧易生物医学科技有限公司
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