分析与检测

食品中羊肉源性成分微滴数字PCR定量方法的建立

  • 陈传君 ,
  • 金鹭 ,
  • 林华 ,
  • 胡滨 ,
  • 韩国全 ,
  • 陈世界 ,
  • 张婧 ,
  • 安微 ,
  • 杨苗
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  • 1(四川农业大学 食品学院,四川 雅安,625000)
    2(成都海关,四川 成都,610041)
    3(食品安全检测四川省重点实验室,四川 成都,610041)
硕士研究生(胡滨副教授和韩国全副教授为共同通讯作者,E-mail:hubin2555@sina.com,153192921@qq.com)

收稿日期: 2019-10-21

  网络出版日期: 2020-04-24

基金资助

国家海关总署科技计划项目(动物源性成分定量检测技术及在非法添加打假执法中的应用研究(2017IK019))

Quantification of mutton-derived ingredients in food by droplet digital PCR

  • CHEN Chuanjun ,
  • JIN Lu ,
  • LIN Hua ,
  • HU Bin ,
  • HAN Guoquan ,
  • CHEN Shijie ,
  • ZHANG Jing ,
  • AN Wei ,
  • YANG Miao
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  • 1 (College of food, Sichuan Agriculture University, Ya'an 625000, China)
    2 (Chengdu Customs, Chengdu 610041, China)
    3 (Food Safety Detection Key Laboratory of Sichuan,Chengdu 610041, China)

Received date: 2019-10-21

  Online published: 2020-04-24

摘要

为了对肉制品中羊肉源性成分进行准确定量,该研究采用微滴数字PCR(ddPCR)技术对羊肉的单拷贝基因进行定量检测,根据基因拷贝数建立肉制品中羊肉源性成分ddPCR的定量检测方法。结果表明,通过向样品中添加牛肉作为内标,建立了基于DNA拷贝数与样品质量间线性关系对羊肉源性成分进行ddPCR内标定量的方法,实现了从靶基因拷贝数到样品质量间的一步转化。该方法在检测出0.01%的羊肉源性成分时,检测结果达0.12 copies/μL,能够对含量5%以上的羊肉源性成分进行准确定量。通过对已知成分的混合样品和市售样品进行检测,显示该方法能够准确检出不同样品中羊肉源性成分含量。因此,该方法在肉及肉制品中羊肉源性成分检测和掺假鉴别方面具有较大应用潜力。

本文引用格式

陈传君 , 金鹭 , 林华 , 胡滨 , 韩国全 , 陈世界 , 张婧 , 安微 , 杨苗 . 食品中羊肉源性成分微滴数字PCR定量方法的建立[J]. 食品与发酵工业, 2020 , 46(6) : 229 -237 . DOI: 10.13995/j.cnki.11-1802/ts.022599

Abstract

In order to accurately quantify the proportion of mutton-derived ingredients in meat products, the single-copy gene of mutton for quantitatively detection of mutton ingredients by the droplet digital polymerase chain reaction (ddPCR) was established in meat products. The result showed that an internal standard for quantitative analysis of the proportion of mutton-derived ingredients with ddPCR was established based on the linear relationship between DNA copy number and sample quality by adding beef as internal standard. The method developed here realized one-step transformation from target gene copy number to sample quality. The detection limit reached 0.01% of sheep-source ingredients which equaled to 0.12 copies/μL. In reality, it could accurately quantify the mutton-derived ingredients which was more than 5% of mutton in meat products. The quantitative analysis of commercial samples showed that ddPCR could accurately determine the proportion of mutton-derived ingredients in meat samples. So ddPCR had great potential in the detection and adulteration identification of mutton-derived ingredients in meat and meat products.

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