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Ingredients of animal origins in meat products identified by using mass spectrometry

An animal-derived ingredient, animal-derived technology, applied in the field of test kits, can solve the problems of sensory properties, protein component damage, inconvenience in meat product trading and import, and difficulty in accurate identification of trace components, and achieves low false positives and detection results. Accurate and stable, highly automated effects

Active Publication Date: 2014-05-21
BIOYONG TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, for processed cooked meat products, due to the destruction of their sensory properties, protein components and other immunogenic substances, it is difficult to carry out accurate identification of meat varieties by traditional methods, which brings great challenges to the trade and import of meat products. great inconvenience
[0006] Chinese patent application 200710175520.6, "Chicken, beef, pork and horse meat multiplex PCR rapid detection kit and its application", this method can quickly identify animal-derived components, but it is difficult to accurately identify trace components in meat products, and requires The result of electrophoresis gel judgment is not economical and practical
This method requires a lot of gel electrophoresis to analyze its gel pattern. The whole process is relatively cumbersome and still stays in the traditional PCR detection technology.

Method used

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  • Ingredients of animal origins in meat products identified by using mass spectrometry
  • Ingredients of animal origins in meat products identified by using mass spectrometry
  • Ingredients of animal origins in meat products identified by using mass spectrometry

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Example 1: Primer design and synthesis.

[0064] Design universal PCR primer core sequences (SEQ ID No: 1 to SEQ ID No: 2) and specific extension primer core sequences (SEQ ID No: 3 to SEQ ID No: 8) for pigs, cattle, sheep, chickens, ducks, and rabbits ).

[0065] Among them, in order to prevent PCR primers from entering the detection window of the mass spectrometer and interfering with the detection effect, a certain number of bases can be added to the core sequence (SEQ ID No: 1 to SEQ ID No: 2) at the 5' end of each PCR primer , such as a 10bp tag (ACGTTGGATG), to increase the molecular weight of the PCR primers, thereby exceeding the detection window of the mass spectrometer.

[0066] Relevant primers were synthesized at Sangon Bioengineering (Shanghai) Co., Ltd.

[0067] The following detection process is performed according to the "Meat Product Identification Kit (Time-of-Flight Mass Spectrometry) Instructions" (hereinafter referred to as "Instructions") issued ...

Embodiment 2

[0068] Example 2: Extraction of genomic DNA in standard meat products.

[0069] Buy one piece of pig, cow, sheep, chicken, duck, and rabbit adult tissue (such as front leg or hind leg) in the market, and select an appropriate amount of muscle tissue inside the tissue. Genomic DNA in muscle tissue was extracted using a commercial nucleic acid extraction kit (such as DNeasy Blood and Tissue kit from QIAGEN). To extract DNA from mixed meat, the steps are as follows:

[0070] A. Take 4g of mixed meat, grind it into meat powder at room temperature, put the meat powder in a 10ml centrifuge tube, add 1ml of STE liquid, 1ml of 10% SDS liquid by volume, 150μl of 20mg / ml proteinase K, Water bath at 56°C for 1-3h;

[0071] B. Add it into 5ml of phenol-chloroform-isoamyl alcohol mixture, mix well, and centrifuge for 4min with a 10000rpm centrifuge; wherein the volume ratio of phenol-chloroform-isoamyl alcohol mixture is phenol 25: chloroform 24: isoamyl alcohol amyl alcohol 1;

[0072...

Embodiment 3

[0077] Embodiment three: standard biological experiment.

[0078] Using ABI9700 PCR instrument, the extracted genomic DNA was tested according to the instructions.

[0079] The components used in the kit for PCR, PCR product purification and single base extension are:

[0080] serial number

component name

main ingredient

Specification

1

PCR mix

dNTPs, MgCl2, PCR primers

360ul / tube x1 tube

2

PCR enzyme

Taq enzyme

24ul / tube x1 tube

3

SAP Enzyme Mix

SAP enzyme

24ul / tube x1 tube

4

Extension Primer Mix

extension primer

24ul / tube x1 tube

5

elongase mix

iPLEX enzymes, ddNTPs

24ul / tube x1 tube

[0081] According to the manual, the specific operation method is as follows:

[0082] 1. PCR amplification

[0083] 1.1 In the PCR preparation area, prepare 200ul PCR reaction tubes according to the number of samples to be tested (including positive quality control prod...

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PUM

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Abstract

The invention discloses a primer system for detecting animal origins in meat products. The primer system comprises a pair of amplification primers and six extending primers, wherein the sequences of the pair of amplification primers are selected from SEQ ID No:1-2, and the sequences of the extending primers are respectively selected from SEQ ID No:3-8. A product prepared based on the primer system can be used for identifying the ingredients of pig, cattle, sheep, chicken, duck and rabbit origins in meat products. By using the product, the identification on sources of meat products can be implemented, therefore, the product has an important role in the aspects of fighting against food adulteration and forging, checking food labels, ensuring the right of information of consumers, and the like.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a detection method and product for identifying animal-derived components in meat products. Methods and corresponding kits for detecting components derived from chicken, duck and rabbit. Background technique [0002] Meat products are an important food in the human diet, which contain high-quality protein and a variety of essential amino acids, and have high nutritional value. With the continuous change of people's dietary structure, the demand for meat products has increased, and there have been many shoddy and counterfeit beef and mutton phenomena in the market. For example, it is common to use chicken, duck and pork as mutton, and choose pig and rabbit Passing off as beef, which seriously damages the financial and health of consumers. [0003] In addition, due to the incidence and prevalence of diseases such as mad cow disease, foot-and-mouth disease of pigs and cattle, and bird fl...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/686C12Q2533/101C12Q2565/627
Inventor 马庆伟张海燕钟逾黎大洪向华林燕刘杰
Owner BIOYONG TECH
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