微滴式数字聚合酶链式反应对香肠制品中鸡、猪、牛源性成分的定量分析  

Quantitative Analysis of Chicken-,Porcine-and Bovine-Derived Ingredients in Sausage Products by Droplet Digital Polymerase Chain Reaction

作　　者：刘立兵 陈敏娜 孙晓霞 张亦琴 付琦 钱云开 周巍 郭春海 王建昌 LIU Libing;CHEN Minna;SUN Xiaoxia;ZHANG Yiqin;FU Qi;QIAN Yunkai;ZHOU Wei;GUO Chunhai;WANG Jianchang(Technology Center of Shijiazhuang Customs,Shijiazhuang 050051,China;Logistics Management Center of Shijiazhuang Customs,Shijiazhuang 050051,China;Technology Center of Qinhuangdao Customs,Qinhuangdao 066002,China;Hebei Food Inspection and Research Institute,Shijiazhuang 050200,China)

机构地区：[1]石家庄海关技术中心,河北石家庄050051 [2]石家庄海关后勤管理中心,河北石家庄050051 [3]秦皇岛海关技术中心,河北秦皇岛066002 [4]河北省食品检验研究院,河北石家庄050200

出　　处：《肉类研究》2020年第8期51-56,共6页Meat Research

基　　金：河北省科技计划项目(17275507D)。

摘　　要：为实现肉制品中动物源性成分的定量检测,基于微滴式数字聚合酶链式反应(droplet digital polymerase chain reaction,ddPCR),以鸡的转化生长因子β-3基因、猪的朊蛋白基因和牛的生长激素基因为靶基因,建立香肠制品中鸡、猪、牛源性成分的定量检测方法。结果表明:所建立ddPCR方法特异性强,能特异性检测相应的鸡、猪、牛源性成分;根据肉粉质量(mg)与DNA质量浓度(ng/μL)、DNA质量浓度(ng/μL)与基因拷贝数浓度(copies/μL)之间的线性关系,得到鸡肉粉、猪肉粉和牛肉粉质量(x1)与基因拷贝数浓度(y 2)之间的关系式为:鸡:x1=n×y2/273.946-n/886.557+0.216;猪:x1=n×y2/64.950+n/60.644+0.215;牛:x1=n×y2/62.839+n/12.646+1.218(n为稀释倍数);基于建立的ddPCR方法对64份市售不同种类香肠样品进行检测,在1份原料标识仅为“猪肉”的香肠中检出鸡源性成分含量为18.68%。本研究建立的ddPCR方法能够实现香肠中鸡、猪、牛源性成分的准确定量检测,并可以此判断“蓄意掺假”或“无意带入”。A droplet digital polymerase chain reaction(ddPCR) assay based on the transforming growth factor beta 3 gene of chicken, the porcine prion protein gene and the bovine growth hormone gene as the target genes were developed for the quantitative detection of chicken-, porcine-and bovine-derived ingredients in sausage products. The results showed that the established assay was highly specific for the detection of chicken-, porcine-and bovine-derived ingredients, but not for other tested animal components. Based on the linear relationship of DNA concentration(ng/μL) with meat mass(mg) and the number of gene copies per unit volume(copies/μL), the linear equations between meat mass(x1) and the number of n × y gene copies(y2) were established as follows: x1=n×y2/273.946-n/886.557+0.216 for chicken;x1=n×y2/64.950+n/60.644+0.215 for pork;and x1=n×y2/62.839+n/12.646+1.218 for beef(where n is dilution factor). The ddPCR assay was used for the quantitative detection of 64 samples of different kinds of comercially available sausages. It was found that one of these samples, labelled as containing only pork, contained 18.68% chicken-derived ingredients. The ddPCR assay could allow accurate and quantitative analysis of chicken-, porcine-and bovine-derived ingredients in sausage, and could be used to discriminate intentional and unintentional adulteration.

关 键 词：微滴式数字聚合酶链式反应 鸡、猪、牛源性成分 定量分析 肉类掺假 

分 类 号：TS207.3[轻工技术与工程—食品科学]