肉制品中牛源性成分芯片数字PCR定量检测方法的建立
作者:
作者单位:

河南省食品和盐业检验技术研究院 国家市场监管重点实验室(食品安全快速检测与智慧监管技术), 河南 郑州 450003

作者简介:

邓迎春 女 高级工程师 研究方向为食品检验 E-mail:dengyingchun@126.com

通讯作者:

陶健 男 正高级工程师 研究方向为食品检验 E-mail:369694930@qq.com

中图分类号:

R155

基金项目:

河南省市场局科技项目(2022sj21);河南省科技计划项目(222102410011)


Quantitative detection methods of bovine-derived ingredients in meat product by chip digital PCR
Author:
Affiliation:

He’nan Food and Salt Industry Inspection Technology Research InstituteKey Laboratory of Food Safety Quick Testing and Smart Supervision Technology for State Market Regulation, He’nan Zhengzhou 450003, China

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    摘要:

    目的 为了对肉制品中牛肉源性成分进行准确定量,本文采用数字PCR(dPCR)技术对牛肉的线粒体Cytb基因进行定量检测,根据基因拷贝数建立了肉制品中牛肉源性成分PCR的定量检测方法。方法 参照标准设计的牛源性特异性引物、探针,猪源性特异性引物、探针,鸭源性特异性引物、探针以及动物源性成分通用引物、探针,建立了双重数字PCR体系。结果 在一定范围内生鲜牛肉质量与DNA含量、DNA含量与DNA拷贝数之间均呈现明显的线性关系,并以DNA含量为中间值计算出DNA拷贝数(C)与生鲜牛肉质量之间的换算公式:M=0.020 9C+0.676。应用建立的dPCR方法对构建的肉样模型进行检测,结果显示牛肉定量检测值与实际质量基本一致,且受外源物种的干扰小。运用该方法对抽取的10份市售牛肉样品检测,同时通过特异/通用引物扩增的拷贝数之比(%)辅助判断肉制品中是否存在非牛源性的其他动物源性成分掺假,检测出部分样品不符合标签规定。结论 该方法可实现对牛源性成分的量化检测,能够作为区分故意添加和无意污染的依据,为执法监管部门提供有力的技术保障。

    Abstract:

    Objective In order to accurately quantify the proportion of bovine-derived ingredients in meat products, the Mitochondrial Cytb gene of bovine for quantitatively detection of bovine ingredients by the digital PCR (Dpcr) technology was developed in this paper.Methods This duplex dignal PCR was developed and optimized by designing bovine-derived specific, duck-derived specific, porcine-derived specific and animal-derived universal primers and probes according to standards.Results Good linear relationships between the raw beef meat weight and DNA content and between the DNA content and DNA copy number. By using the dPCR method, we found good linear relationships between the raw meat weight and DNA content and between the bovine -derived DNA content and DNA copy number. Using the DNA content as an intermediate value, we established the following formulae for calculating the weight of the original raw beef meat from the specific DNA copy number: Mbeef=0.020 9C+0.676. The mixed samples of known bovine quality were tested, and the results showed that the method can accurately quantify bovine-derive dingredients, which has litter interference by foreign species. Using this method to test 10 samples of the bovine-derived products on the market, by using the ratio of universal and specific amplified copy number, it could determine whether other animal-derived components were present in the beef meat product. it was found that the content of some samples was not up to standard.Conclusion This method can realize the quantitative detection of bovine origin components, which can be used as a basis to distinguish intentional addition from unintentional pollution and provide a strong technical guarantee for law enforcement and supervision departments.

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邓迎春,郭旭光,苏利霞,陶健.肉制品中牛源性成分芯片数字PCR定量检测方法的建立[J].中国食品卫生杂志,2024,36(5):533-540.

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  • 收稿日期:2023-05-06
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  • 在线发布日期: 2024-08-09
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