摘要
该文建立黄鱼(大黄鱼、小黄鱼)鱼源性成分鉴别方法,针对大黄鱼、小黄鱼线粒体细胞色素b(Cyt b)保守序列,设计引物和Taqman荧光探针,优化反应体系与参数,分别建立了多重实时荧光定量聚合酶链式反应(polymerase chain reaction,PCR)检测方法和微滴式数字PCR检测方法。两种方法均能有效识别大小黄鱼鱼源性成分,所建立的多重实时荧光定量PCR检测方法不仅同时检测大、小黄鱼目标基因片段,而且通过监测内参基因扩增情况判断核酸提取效果。数字PCR检测法可以直接显示扩增体系中阴性、阳性微滴的数量,在作出定性的同时,有助于了解样本中各组分组成比例情况。
To establish methods for identifying derived components of yellow croaker(including Larimichthys crocea and Larimichthys polyactis).The primers and Taqman fluorescent probes were designed for the conserved sequence of mitochondrial cytochrome b(Cyt b)of L.crocea and L.polyactis,and the reaction system and parameters were optimized.Multiple real-time fluorescent quantitative PCR detection methods and micro drop digital PCR detection methods were established respectively.Both methods effectively identified the derived components of L.crocea and L.polyactis.The multiplex real-time fluorescent quantitative PCR detection method not only detected the target gene fragments of L.crocea and L.polyactis at the same time but also judged the nucleic acid extraction effect by monitoring the amplification of internal reference genes.Digital PCR detection directly displayed the number of negative and positive droplets in the amplification system,which helped understand the composition proportion of each component in the sample while making qualitative determination.
作者
邵彪
陈旭
蔡茜茜
高利亭
徐陈红
张霞
陈刚
SHAO Biao;CHEN Xu;CAI Xixi;GAO Liting;XU Chenhong;ZHANG Xia;CHEN Gang(Nantong Products Quality Supervision and Inspection Institute,Nantong 226011,Jiangsu,China;Nantong Key Laboratory of Food Hazards,Nantong 226011,Jiangsu,China;College of Biological Science and Engineering,Fuzhou University,Fuzhou 350108,Fujian,China)
出处
《食品研究与开发》
CAS
2024年第14期142-147,共6页
Food Research and Development
基金
南通市科技项目(JC2020108)
江苏省市场监督管理局科技计划项目(KJ204137)。
关键词
大黄鱼
小黄鱼
鉴别
实时荧光定量PCR
数字PCR
Larimichthys crocea
Larimichthys polyactis
identification
real-time fluorescent quantitative PCR
digital PCR
作者简介
邵彪(1983-),男(汉),正高级工程师,博士,研究方向:食品分析检验。
