双重实时荧光PCR定量检测动物产品中牛源性成分

doi:10.13560/j.cnki.biotech.bull.1985.2017-1129

生物技术通报 ›› 2018, Vol. 34 ›› Issue (9): 190-194.doi: 10.13560/j.cnki.biotech.bull.1985.2017-1129

双重实时荧光PCR定量检测动物产品中牛源性成分

高志强, 汪琳, 蒲静, 尹羿, 张伟, 赵相鹏, 姚震宇

北京出入境检验检疫局技术中心,北京 100026

收稿日期:2017-12-27 出版日期:2018-09-26 发布日期:2018-10-10
作者简介:高志强,男,博士,研究方向：动物检验检疫;E-mail：gaozhq@bjciq.gov.cn
基金资助:
国家“十三五”重点研发计划项目（2017YFF0210305）

Duplex Real-time PCR Methods for Quantitative Detection of Bovine Derived Materials in Animal Products

GAO Zhi-qiang, WANG Lin, PU Jing, YIN Yi, ZHANG Wei, ZHAO Xiang-peng, YAO Zhen-yu

Beijing Entry-Exit Inspection and Quarantine Bureau Technical Center,Beijing 100026

Received:2017-12-27 Published:2018-09-26 Online:2018-10-10

摘要/Abstract

摘要： 旨在建立一种定量检测动物产品中牛源性成分含量的双重实时荧光PCR方法。以牛卫星IV DNA为检测靶基因,以肌肉生长抑制素基因作为内对照合成引物探针,采用基于TaqMan的双重实时荧光PCR技术对已知牛肉添加比例的冻干混合肉粉标准品进行检测并建立线性模型,并应用模拟样品和送检样品进行了验证检测。结果显示,该方法可检出牛源成分含量为0.1%的冻干肉粉,不与其他种动物组织发生交叉反应,重复性试验的相对标准偏差小于5%,方法的扩增效率为93.6%。该方法适用于肉品中牛源性成分的定量检测,可用于测定市场上动物产品中的牛源性成分含量。

关键词: 双重实时荧光PCR, 定量检测, 牛源性成分

Abstract: The objective of this work is to develop a duplex real-time quantitative PCR method for detecting the amount of bovine derived material in animal product. Using bovine satellite IV DNA as detection target gene and myostatin gene as an internal control to synthesize primer probe,the standard freeze-dried meat mixture powder with known beef proportion was detected by duplex real-time PCR based on TaqMan,and the linear model was established. Then the model was verified by detecting mocked and delivered samples. The method detected 0.1% bovine material in freeze-dried meat powder,and no cross reaction with the tissues of other animal species occurred. The relative standard deviation of repeatability test was < 5%,and the amplification efficiency of the method was 93.6%. Conclusively,the established method is suitable for quantitative detection of bovine derived material in meat products,and can be used to determine the content of bovine derived material in animal products on the market.

Key words: duplex real-time PCR, quantitative detection, bovine derived material

高志强, 汪琳, 蒲静, 尹羿, 张伟, 赵相鹏, 姚震宇. 双重实时荧光PCR定量检测动物产品中牛源性成分[J]. 生物技术通报, 2018, 34(9): 190-194.

GAO Zhi-qiang, WANG Lin, PU Jing, YIN Yi, ZHANG Wei, ZHAO Xiang-peng, YAO Zhen-yu. Duplex Real-time PCR Methods for Quantitative Detection of Bovine Derived Materials in Animal Products[J]. Biotechnology Bulletin, 2018, 34(9): 190-194.

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双重实时荧光PCR定量检测动物产品中牛源性成分

Duplex Real-time PCR Methods for Quantitative Detection of Bovine Derived Materials in Animal Products

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