基于微滴式数字聚合酶链式反应技术的肉制品中鸭源性成分的定量检测  

Quantitative Detection of Duck-Derived Ingredients in Meat Products by Droplet Digital Polymerase Chain Reaction

作　　者：刘立兵 员丽娟 陈敏娜 王金凤 付琦 孙晓霞 王建昌 LIU Libing;YUAN Lijuan;CHEN Minna;WANG Jinfeng;FU Qi;SUN Xiaoxia;WANG Jianchang(Technology Center of Shijiazhuang Customs,Shijiazhuang 050051,China;Technology Center of Xi'an Customs,Xi'an 710075,China)

机构地区：[1]石家庄海关技术中心,河北石家庄050051 [2]西安海关技术中心,陕西西安710075

出　　处：《肉类研究》2021年第3期30-32,33,34,共5页Meat Research

基　　金：“十三五”国家重点研发计划重点专项(2020YFF0305000)。

摘　　要：为实现肉制品中鸭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)技术,以鸭生长激素(growth hormone,GH)基因为靶基因,建立肉制品中鸭源性成分的定量检测方法。结果表明:所建立的ddPCR方法能特异性检测鸭源性成分,灵敏性为11.1 copies/μL;根据鸭肉粉质量(mg)与DNA质量浓度(ng/μL)、DNA质量浓度(ng/μL)与鸭GH基因拷贝数浓度(copies/μL)之间的线性关系,建立鸭肉粉质量(x)与鸭GH基因拷贝数浓度(y)之间的关系式为x=(n×y)/725.046-n/1317.06+0.153;模拟混合样本中鸭肉粉质量分数5%-100%时,鸭源性成分检测的绝对误差均小于±1.52%,变异系数均小于10.38%,回收率为98.35%~125.32%;在88份商品化肉制品中,11份肉制品中检出鸭源性成分,检出率为12.5%,且鸭源性成分含量为18.6%~87.4%。本研究所建立的鸭源性成分ddPCR方法有较好的准确性和较强的适用性,能够应用于肉制品中鸭源性成分的定量检测。A droplet digital polymerase chain reaction assay(ddPCR)based on the duck growth hormone(GH)gene as the target gene was developed for the quantitative detection of duck-derived ingredients in meat products.The results showed that the assay was highly specific for the detection of duck derived ingredients,but not the other tested animal components,and its limit of detection was 11.1 copies/μL.According to the linear relationship between the mass of duck meat(mg)and DNA concentration(ng/μL),and that between DNA concentration(ng/μL)and the number of GH gene copies(copies/μL),the linear equation between the mass of duck meat(x)and the number of GH gene copies(y)was established as follows:x=(n×y)/725.046-n/1317.06+0.153.When the percentage of duck meat powder ranged from 5%to 100%in simulated mixed samples,the absolute error was less than±1.52%,and the coefficient of variation was less than 10.38%with a recovery between 98.35%and 125.32%.Duck-derived ingredients were detected at levels of 18.6%-87.4%in 11 out of88 samples of commercial meat products,with a detection rate of 12.5%.The developed ddPCR assay demonstrated good accuracy and applicability for the quantitative determination of duck-derived component in commercial meat products.

关 键 词：肉制品 鸭源性成分 生长激素基因 微滴式数字聚合酶链式反应 定量检测 

分 类 号：TS207.3[轻工技术与工程—食品科学]