羊口疮病毒和丝状支原体簇双重PCR检测方法的建立及应用

发布时间：2018-03-21 02:02

本文选题：羊口疮病毒　切入点：丝状支原体簇　出处：《中国预防兽医学报》2017年04期 　论文类型：期刊论文

【摘要】：为建立一种同时快速检测羊口疮病毒(ORFV)和丝状支原体簇(Mm cluster)的方法,本研究根据ORFV的B2L基因和Mm cluster的16S r RNA基因序列,设计合成了2对特异性引物,通过条件优化,建立了同时检测ORFV核酸和Mm cluster核酸的双重PCR方法。该方法可以同时扩增出ORFV和Mm cluster的特异性片段,而对其它常见病原的DNA模板均无扩增,对ORFV和Mm cluster的最低检出量分别为3.8×10~4拷贝/μL和1.3×10~3拷贝/μL。利用该方法对36份临床样品进行检测,结果显示双重PCR检测结果与单一PCR检测结果符合率为100%。本研究建立的双重PCR方法特异性强,敏感性高,可重复性好,为临床ORFV和Mm cluster的快速检测、诊断及流行病学调查提供了方法。
[Abstract]:In order to establish a method for rapid detection of sheep mouth sore virus (ORFV) and mycoplasma mycoplasma (Mm cluster) simultaneously, two pairs of specific primers were designed and synthesized according to the sequence of B2L gene of ORFV and 16s r RNA gene of Mm cluster. A double PCR method for simultaneous detection of ORFV nucleic acid and Mm cluster nucleic acid was established. The specific fragments of ORFV and Mm cluster could be amplified simultaneously by this method, but the DNA templates of other common pathogens were not amplified. The minimum detectable amounts of ORFV and Mm cluster were 3.8 脳 10 ~ 4 copies / 渭 L and 1.3 脳 10 ~ 3 copies / 渭 L respectively. 36 clinical samples were detected by this method. The results showed that the coincidence rate between double PCR and single PCR was 100%. The double PCR method established in this study has the advantages of high specificity, high sensitivity and good repeatability. It is a rapid detection method for clinical ORFV and Mm cluster. Diagnostic and epidemiological surveys provide a method.
【作者单位】：福建省农业科学院畜牧兽医研究所;
【基金】：福建省公益类科研院所专项(2014R1023-6) 福建省公益科研院所专项(2014R1023-15) 福建省科技创新平台建设项目(2014N2003-5) 国家重点研发计划项目(2016YFD0500906)
【分类号】：S852.6

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