基于实时荧光定量技术建立定量检测乳粉中羊源性成分的方法  

Establishment of a Fluorescence-Based Quantitative Real-Time PCR Method for Detection for Caprine-Derived Ingredients in Milk Powder

作　　者：陈晨 张瑞 王玉欣 张同贺 张子仑 贾文珅[2] 周巍 CHEN Chen;ZHANG Rui;WANG Yuxin;ZHANG Tonghe;ZHANG Zilun;JIA Wenshen;ZHOU Wei(Hebei Food Safety Key Laboratory,Key Laboratory of Special Food Supervision Technology for State Market Regulation,Hebei Engineering Research Center for Special Food Safety and Health,Hebei Food Inspection and Research Institute,Shijiazhuang 050227,China;Beijing Academy of Agricultural and Forestry Sciences,Beijing 100050,China)

机构地区：[1]河北省食品检验研究院,河北省食品安全重点实验室,国家市场监管重点实验室(特殊食品监管技术),特殊食品安全与健康河北省工程研究中心,河北石家庄050227 [2]北京市农林科学院,北京100050

出　　处：《乳业科学与技术》2023年第6期20-24,共5页JOURNAL OF DAIRY SCIENCE AND TECHNOLOGY

基　　金：河北省市场监督管理局科研计划项目(2022ZD11)。

摘　　要：基于实时荧光定量聚合酶链式反应建立了乳粉中羊源性成分相对定量检测方法。对羊引物进行特异性、灵敏度和稳定性实验。通过模拟不同质量浓度羊乳粉与马乳粉混合样本,根据其ΔCt值的函数关系进行线性拟合进而绘制标准曲线,建立乳粉中羊源性成分相对定量检测。结果显示,该方法最低检测限为0.000 1 ng/μL、标准曲线方程为y=0.720 9x+5.651 9(R2=0.987 5)、回收率为96.22%~112.00%、组间变异系数≤0.79%、组内变异系数≤1.77%。This study established a fluorescence-based quantitative real-time polymerase chain reaction method for the detection of caprine-derived ingredients in milk powder.The specificity,sensitivity,and stability of the caprine-specific primers were evaluated.By linear fitting of the difference in cycle threshold(ΔCt)as a function of the mixing proportion between goat and horse milk powder,a calibration curve for the relative quantitation of caprine-derived ingredients in milk powder was established as follows:y=0.7209x+5.6519(R²=0.9875),and the minimum detection limit of this method was 0.0001 ng/μL.The recoveries of the proposed method were 96.22%-112.00%,and the inter-and intra-group coefficient of variation was≤0.79%and≤1.77%,respectively.

关 键 词：羊乳粉 马乳粉 实时荧光定量聚合酶链式反应 掺假检测 

分 类 号：TS251.7[轻工技术与工程—农产品加工及贮藏工程]