实时荧光PCR法快速鉴别狐狸貉子肉源性成分研究

Rapid Detection of Alopex lagopus- and Nyctereutes procyonoides-derived Constituents of Meat Products Using RT-PCR

摘要:

根据狐狸线粒体基因组中的保守序列和貉子线粒体D-loop基因保守序列设计狐狸、貉子特异性引物和TaqMan探针，建立一种基于实时荧光聚合酶链式反应的肉及肉制品狐狸、貉子源性成分测定方法，通过特异性、灵敏性、线性检测对该方法体系进行检验和评价。本研究建立的狐狸、貉子源性成分实时荧光聚合酶链式测定方法体系具有良好的特异性及灵敏性，最低可检测0.5 pg/μL纯狐狸DNA和5 pg/μL纯貉子DNA。本研究建立的狐狸貉子源性成分荧光PCR检测方法，可以用来检测实际样品中是否含有狐狸貉子源性。

Abstract:

Alopex lagopus- and Nyctereutes procyonoides-specific primers and TaqMan probes were designed based on the conserved sequences in the mitochondrial genome of the Alopex lagopus and the displacement-loop (D-loop) region of Nyctereutes procyonoides. A real-time polymerase chain reaction (RT-PCR)-based method for the determination of Alopex lagopus- and Nyctereutes procyonoides-derived constituents of commercial meat products was developed in this study. The established RT-PCR method was evaluated for specificity, sensitivity, and linearity; the method showed good specificity and sensitivity, and the limits of detection for Alopex lagopus DNA and Nyctereutes procyonoides DNA were 0.5 pg/μL and 5 pg/μL, respectively. In summary, this RT-PCR method can be used for the identification of Alopex lagopus- and Nyctereutes procyonoides-derived constituents in commercial samples.