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中国精品科技期刊2020
张秀平, 苗丽. 实时荧光PCR定量测定肉制品中羊源性成分[J]. 食品工业科技, 2019, 40(7): 119-123,130. DOI: 10.13386/j.issn1002-0306.2019.07.021
引用本文: 张秀平, 苗丽. 实时荧光PCR定量测定肉制品中羊源性成分[J]. 食品工业科技, 2019, 40(7): 119-123,130. DOI: 10.13386/j.issn1002-0306.2019.07.021
ZHANG Xiu-ping, MIAO Li. Quantitative Detection of Ovine-derived Materials in Meat Products by Real-time Fluorescent PCR[J]. Science and Technology of Food Industry, 2019, 40(7): 119-123,130. DOI: 10.13386/j.issn1002-0306.2019.07.021
Citation: ZHANG Xiu-ping, MIAO Li. Quantitative Detection of Ovine-derived Materials in Meat Products by Real-time Fluorescent PCR[J]. Science and Technology of Food Industry, 2019, 40(7): 119-123,130. DOI: 10.13386/j.issn1002-0306.2019.07.021

实时荧光PCR定量测定肉制品中羊源性成分

Quantitative Detection of Ovine-derived Materials in Meat Products by Real-time Fluorescent PCR

  • 摘要: 为准确定量肉制品中羊源性成分的含量,以羊单拷贝基因为扩增靶基因,设计合成羊源的引物、探针,制备羊源和脊椎动物通用基因的重组质粒以构建标准曲线,通过方程算出各自的拷贝数,根据拷贝数的比值计算样品中羊肉源成分占总肉成分的百分比含量,建立荧光定量PCR检测体系,并对已知目标肉含量的混合肉样进行定量检测。结果显示,该方法具有良好的特异性和灵敏度,最低检测到羊的DNA含量为0.02 ng/μL。对已知目标肉含量的混合肉样进行定量检测发现,羊肉掺入量为90%、50%时所得结果相对准确,相对标准偏差分别为5.69%和10.82%, 且不受外源物种的干扰。本试验建立的定量PCR方法能够准确检测出不同样品中羊肉的含量,并发现存在掺假现象。

     

    Abstract: In order to determine the contents of ovine in meat products,primers and probes were designed with their conservative regions of single copy gene to prepare recombinant plasmids for the universal gene of sheep and vertebrate to construct a standard curve. A fluorescent quantitative PCR detection system was established by calculating the number of copies by the equation,calculating the percentage contents of the mutton source in the sample based on the ratio of the copy number and the mixed meat samples with the known target meat content were quantitatively detected. The results showed that the PCR method had good specificity and sensitivity,and the minimum DNA contents of ovine was 0.02 ng/μL. Quantitative detection with a known target meat showed that the results were relatively accurate when the incorporation was 90% and 50%,and the relative standard deviations were 5.69% and 10.82%, respectively. These results were not interfered by exogenous species. The quantitative PCR method established in this experiment could accurately detect the contents of mutton in different samples and find the adulteration.

     

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