食品科学

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微滴数字PCR法对肉制品中牛源和猪源成分的定量分析

苗 丽1,张秀平2,陈 静2,李 轲2,王永杰2,白 杰2   

  1. 1.河南出入境检验检疫局,河南 郑州 450003;2.河南检验检疫鉴定咨询中心,河南 郑州 450003
  • 出版日期:2016-04-25 发布日期:2016-04-13

Quantitative Analysis of Bovine and Porcine Ingredients in Meat Products by Droplet Digital PCR

MIAO Li1, ZHANG Xiuping2, CHEN Jing2, LI Ke2, WANG Yongjie2, BAI Jie2   

  1. 1. Henan Entry-Exit Inspection and Quarantine Bureau, Zhengzhou 450003, China;
    2. Identification Consulting Center of Henan Inspection and Quarantine, Zhengzhou 450003, China
  • Online:2016-04-25 Published:2016-04-13

摘要:

为准确检测肉及肉制品中肉源成分的含量,实验基于微滴数字聚合酶链式反应(droplet digital polymerasechain reaction,ddPCR)技术,建立了定量检测肉及肉制品中牛肉和猪肉含量的方法。由ddPCR结果可知,在一定范围内生鲜肉质量与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)与生鲜肉质量之间的换算公式M牛=0.062C-0.943,M猪=0.045C-1.72。应用建立的数字ddPCR方法对已知目标肉种含量的混合肉样进行检测,结果表明测量值和真实值基本一致,且不受外源物种的干扰。通过对市售样品的检测,能够准确检测出不同样品中牛肉和猪肉的含量,并发现存在掺假现象,说明该检测方法具有良好的市场应用前景。本实验建立的ddPCR方法在肉及肉制品中牛肉和猪肉含量的定量检测方面具有较大的应用潜力,可为肉制品真伪鉴别日常检测提供有力的科学依据。

关键词: 微滴数字PCR, 肉制品, DNA拷贝数, 掺杂使假

Abstract:

A highly precise quantitative method based on the droplet digital polymerase chain reaction (ddPCR) technique
was developed to detect bovine and porcine ingredients in meat products. By using the ddPCR method, we found good linear
relationships between the raw meat weight and DNA content and between the 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 meat from the specific DNA copy number: Mbeef = 0.062C - 0.943,Mpork = 0.045C - 1.72. By examining the mixed meat
samples of known compositions, we found that the final quantitative results for the mixed samples were similar to the true
raw meat weights, free from interference from exogenous species. Analysis of commercial samples showed that the ddPCR
quantification system enables determination of the proportions of bovine and porcine ingredients in meat products with good
practicability. Quantitative analysis indicates that ddPCR is highly precise in quantifying beef and pork in meat products and
therefore has the potential to be used in routine analysis and meat adulteration of various species.

Key words: droplet digital polymerase chain reaction, meat products, DNA copy number, meat adulteration

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