荧光定量PCR法检测皮纤维中黄牛、绵羊源性成分研究  

Detection of Cattle and Sheep Derived Materials in Leather Fiberby Double Fluorescence Quantitation PCR

作　　者：周宇东 姚艳玲 孙世元 王文宇 刘骆强 管佳丽 张娜 李慧玲 ZHOU Yudong;YAO Yanling;SUN Shiyuan;WANG Wenyu;LIU Luoqiang;GUAN Jiali;ZHANG Na;LI Huiling(Jiaxing Testing Institute for Food,Drug and Product Quality Control,Jiaxing 314050,China)

机构地区：[1]嘉兴市食品药品与产品质量检验检测院,浙江嘉兴314050

出　　处：《西部皮革》2024年第1期30-33,53,共5页West Leather

摘　　要：为实现皮纤维产品中常见原料黄牛、绵羊源性成分的鉴别与定量分析,采用实时荧光聚合酶链式反应技术比较了58、60℃两个退火温度下,黄牛、绵羊源性成分的扩增效果,利用两种成分浓度比值的对数与两者对应的阈值循环数之差存在线性关系,建立了黄牛、绵羊皮纤维的双重定量PCR(double fluorescence quantitation PCR)检测方法。结果显示,退火温度设置为58℃时,黄牛、绵羊源性成分均能有效扩增,且最低检出的DNA质量浓度均为10^(-3)ng/μL。经荧光定量检测分析,建立标准曲线时,黄牛与绵羊皮纤维源性成分的浓度比值范围为1/9~9/1,标准曲线的相关系数为0.9954。文章建立的黄牛、绵羊皮纤维源性成分的双重定量PCR检测方法,不仅能定性鉴别两种成分,而且能较为准确地进行两者的定量分析,与实际结果偏差为5%。In order to achieve the identification and quantitative analysis of ingredients from cattle and sheep in leather fiber products,real-time fluorescence PCR technology was used to compare the amplification effects of cattle and sheep ingredients at two annealing temperatures of 58 and 60℃.A linear relationship between the logarithm of the concentration ratio of the two components and the difference in their corresponding threshold number of cycles was used to establish a double fluorescence quantification PCR detection method.The results showed that when the annealing temperature was set to 58℃,both cattle and sheep derived components could be effectively amplified,and the lowest detected DNA concentration was 10^(-3) ng/μL.Through fluorescence quantitative detection analysis,when establishing the standard curve,the concentration ratio range between cattle and sheep derived components is 1/9-9/1,and the correlation coefficient of the standard curve is 0.9954.The dual quantitative PCR detection method established in this study for cattle and sheep derived components in leather fiber can not only qualitatively identify the two components,but also accurately perform quantitative analysis of the two,with a deviation of 5%from the actual results.

关 键 词：皮纤维 源性成分 双重荧光定量PCR 

分 类 号：TS57[轻工技术与工程—皮革化学与工程]