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Position: Home > Articles > A RT-PCR Method for Quantitative Determination of Pork-Derived Ingredients in Meat and Meat Products MEAT RESEARCH 2013,27 (12) 11-15

一种基于实时荧光聚合酶链式反应的肉及肉制品中猪源性成分含量测定

作  者:
周彤;李家鹏;田寒友;杨君娜;邹昊;乔晓玲
单  位:
中国肉类食品综合研究中心
关键词:
实时荧光聚合酶链式反应;猪源性成分;定量;肉类掺假
摘  要:
为了建立一种基于实时荧光聚合酶链式反应的肉及肉制品猪源性成分含量测定方法,分别以猪线粒体DNA的NADH4基因和16S rRNA基因为靶位点设计猪特异性引物、探针和通用引物、探针,建立猪源性成分含量测定方法,通过特异性、灵敏性、线性、准确度及市售肉制品检测,对该方法体系进行检验和评价。结果表明:建立的猪源性成分含量实时荧光聚合酶链式反应测定方法体系具有良好的特异性及灵敏性,最低可检测到0.4pg/μL纯猪肉DNA;无论是猪特异性体系还是通用体系都呈现良好的线性关系(R2均达到0.999以上)和较宽的线性范围;通过含猪源性成分的模拟混合肉样检测,样品猪源性成分含量的回收率平均值为122.27%,说明该方法具有较高的准确度;通过市售肉制品检测表明该方法可以用来检测实际样品(包括生鲜样品和熟肉样品)中猪源性成分含量。
译  名:
A RT-PCR Method for Quantitative Determination of Pork-Derived Ingredients in Meat and Meat Products
作  者:
ZHOU Tong;LI Jia-peng;TIAN Han-you;YANG Jun-na;ZOU Hao;QIAO Xiao-ling;China Meat Research Center;
关键词:
real time PCR;;pork-derived ingredients;;quantifi cation;;meat adulteration
摘  要:
This study is aimed to establish a real time PCR(RT-PCR) method for quantitative determination of porkderived ingredients in meat and meat products.The pork-specifi c primers,universal primers and TaqMan probes were designed for the mitochondrial NADH4 gene and 16S rRNA,respectively,and the established RT-PCR method was evaluated by specifi city,sensitivity,linear relationship and accuracy by using it to detect commercial meat products.The method showed a good specifi city and was able to detect 0.4 pg/μL pork DNA.Both pork-specifi c and universal systems showed a good linear relationship(R2 > 0.999) and a wide linear range.The PCR method showed a high accuracy and the average recovery of pork-derived ingredients in meat mixtures was 122.27%.It can be used to test both raw and processed pork products.

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