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Detection technology and method for meat containing animal-derived components

An animal-derived ingredient and animal-derived technology, applied in the field of biotechnology and food safety testing

Inactive Publication Date: 2016-11-23
SHANDONG INST FOR FOOD & DRUG CONTROL
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

However, this method is mainly for qualitative, semi-quantitative or quantitative detection, and is not suitable for the determination of meat content of animal-derived ingredients

Method used

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  • Detection technology and method for meat containing animal-derived components
  • Detection technology and method for meat containing animal-derived components
  • Detection technology and method for meat containing animal-derived components

Examples

Experimental program
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Embodiment Construction

[0079] In Example 1 of the present invention, the meat to be tested is raw meat, specifically beef.

[0080] Using the above-mentioned fluorescent quantitative PCR and 3D digital PCR detection methods, extract DNA from different animal-derived meats, and mix the extracted DNA in different ratios (for example: beef and chicken DNA are 1:1, 1:10, 1:100 equal ratio mixture), carry out the identification of the copy number and percentage content of bovine-derived components, and perform the following steps in sequence:

[0081] 1. Meat Sampling Method

[0082] Sampling of meat samples shall be carried out in accordance with GB / T 9695.19-2008. The fascia of the samples shall be removed, shredded, and ground into a homogenate for later use.

[0083] 2. Extraction method of meat DNA

[0084] Use about 2.0 g of the homogenate, and select Qiagen's Qiagen DNeasy mericon Food Kit Cat. No. 69514 to extract the sample DNA. Experimental procedure for sample DNA extraction:

[0085] (1) ...

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Abstract

The invention provides a detection technology and method for meat containing animal-derived components. The detection technology and method is based on 3D digital PCR (polymerase chain reaction) technology and fluorescent quantitative PCR technology and comprises the following steps: 1) DNA of a to-be-detected sample is extracted, and a PCR amplification template is obtained; 2) a PCR system is prepared, and 3D digital PCR and fluorescent quantitative PCR amplification are performed, wherein the PCR system comprises the PCR amplification template, a universal primer pair, a fluorescent probe and the like and adopts a dose-effect curve of a pure animal-derived sample standard as the standard curve; 3) the copy number and the percentage composition of the animal-derived components in the to-be-detected sample are calculated according to fluorescence signals produced through the 3D digital PCR. According to the method, a PCR liquid is subjected to smear processing with the 3D digital PCR technology, so that interference by background DNA and a substrate is reduced greatly, and the sensitivity can be as low as one copy; meanwhile, by means of the standard curve, direct processing of follow-up samples is facilitated, and operation steps are simplified greatly.

Description

technical field [0001] The present invention relates to the field of biotechnology and food safety detection. Specifically, the present invention provides a technology and method for meat detection of animal-derived components based on 3D digital PCR technology and fluorescent quantitative PCR technology. Background technique [0002] With the globalization of the food supply chain and the development of eating habits in different countries, cultures and religions, the problem of food adulteration driven by economic interests is becoming more and more serious. Among them, the problem of meat adulteration with animal-derived ingredients is emerging one after another, and the market chaos is worrying. Therefore, there is an urgent need for an accurate and trace detection method for animal-derived ingredients in meat. [0003] At present, there are many methods used in the identification of meat and meat products, mainly including morphological identification, immunological id...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6851C12Q1/6888C12Q2531/113C12Q2561/101C12Q2545/113
Inventor 石峰杨俊杰姜梦菡王蒙巩丽萍邢晟张磊
Owner SHANDONG INST FOR FOOD & DRUG CONTROL
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