一种驴和马及狐狸源性成分快速检测方法的研究

发布时间：2018-11-05 10:20

【摘要】：实验旨在建立快速检测驴、马和狐狸源性成分的方法。以驴、马和狐狸的线粒体保守序列16S r RNA基因为靶基因,设计通用引物和以不同荧光素标记的特异Taq Man探针,构建与通用引物共用的阳性扩增内标(Internal Amplification Control,IAC)作为PCR监控反应体系。通过优化PCR反应体系和条件,建立能同时检测驴、马和狐狸源性成分的四重实时荧光PCR方法。结果表明:利用驴、马、狐狸、牛、猪、羊、水貂、狗、鸡、鸭、大豆、小麦、玉米等物种检测没有交叉反应,证明该方法特异性高,准确性好。该方法模板DNA检测灵敏度为0.01 ng。本研究建立的驴、马和狐狸源性四重荧光实时PCR检测体系灵敏度高、特异性强,能有效鉴别动物产品中驴、马和狐狸的源性成分,也可用作相关动物皮张真伪的鉴别方法。
[Abstract]:The aim of this study was to establish a rapid method for the detection of donkeys, horses and foxes. Using the mitochondrial conserved 16s r RNA gene of donkey, horse and fox as target gene, a universal primer and a specific Taq Man probe labeled with different fluorescein were designed to construct a positive amplified (Internal Amplification Control, shared with universal primers. IAC) as PCR monitoring reaction system. By optimizing the PCR reaction system and conditions, a quadruple real-time fluorescent PCR method was established for simultaneous detection of donkey, horse and fox origin components. The results showed that there was no cross reaction among donkey, horse, fox, cow, pig, sheep, mink, dog, chicken, duck, soybean, wheat and corn. The sensitivity of template DNA is 0. 01 ng.. The real-time PCR system for the detection of donkey, horse and fox is highly sensitive and specific. It can be used to identify the origin components of donkey, horse and fox in animal products, and can also be used as a method to identify the authenticity and falsity of related animal skin.
【作者单位】：山东省农业科学院生物技术研究中心山东省农业科学院应用生命科学实验室;山东省食品药品检验研究院;山东省农业科学院畜牧兽医研究所山东省畜禽疫病防治与繁育重点实验室;
【基金】：山东省农业重大应用技术创新课题(鲁财农指[2014]38号) 山东省科技厅山东省优秀中青年科学家科研奖励基金计划项目(BS2010SW024) 济南市高校院所自主创新项目(201401264)
【分类号】：S852.23

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