垂体后叶粉中3种动物源性成分多重real-time PCR检测方法的建立及应用
Establishment and application of multiplex real-time PCR method for rapid detection of three animal-derived components in powdered posterior pituitary
邵长春 1潘秀丽 1魏艳芸 1王月玲 1王蕙1
作者信息
- 1. 兰州市食品药品检验检测研究院/国家市场监管重点实验室(食品中农药兽药残留监控),甘肃 兰州 730050
- 折叠
摘要
目的 基于三重实时聚合酶链式反应(real-time polymerase chain reaction,real-time PCR)技术同时快速检测垂体后叶粉中的猪、牛、羊性动物源性成分.方法 采用CTAB法提取样品及对照品DNA,以线粒体基因为靶基因设计并合成三重real-time PCR 的引物探针,并对建立方法进行验证,用于检测垂体后叶粉样品是否与标识一致.结果猪、牛、羊源性成分的最低检测浓度为10 pg/μL,采用建立的方法检测 3 批垂体后叶粉样品,均只检测出猪源性成分.结论 建立的方法可用于同时检测垂体后叶粉中的猪、牛、羊源性成分,可为相关工作提供技术支持.
Abstract
Objective To determine the porcine,bovine and sheep derived components in powdered posterior pituitary by a mul-tiplex real-time polymerase chain reaction(PCR)method.Methods The DNA was extracted by CTAB method.The mitochon-dria genes were used as target genes to design specific primers and Taq Man probes.The developed method was validated and ap-plied to detect whether the animal-derived components in the samples were consistent with those indicated.Results The mini-mum detection concentration was 10 pg/μL of template DNA.The3 batches of powdered posterior pituitary samples were identified by the successfully established method,and only pig source components were detected.Conclusion The multiplex real-time PCR method can be used for the detection of porcine,bovine and sheep derived components in powdered posterior pituitary.
关键词
垂体后叶粉/多重实时聚合酶链反应/种属鉴定Key words
powdered posterior pituitary/multiplex real-time polymerase chain reaction/species identification引用本文复制引用
出版年
2024
NETL
NSTL
万方数据