绵羊痘病毒TaqMan-MGB荧光定量PCR快速检测方法的建立
发布时间：2018-08-16 13:52
【摘要】：为建立快速检测绵羊痘病毒(SPPV)快速检测方法基因的方法,本研究参照SPPV ORF068基因设计并合成一对特异性引物和一条MGB探针,经条件优化,建立了SPPV TaqMan-MGB荧光定量PCR快速检测方法。结果显示,该方法可以特异性检测SPPV,而对羊口疮病毒和猪水疱病病毒等扩增结果均为阴性;而且检出下限为10拷贝/μL;组内组间重复性试验的变异系数均小于3%。上述结果显示本研究建立的TaqMan-MGB qPCR方法特异性强、灵敏度高、重复性好,能够对SPPV进行快速准确的检测。
[Abstract]:In order to establish a rapid detection method for the (SPPV) gene of sheep pox virus, a pair of specific primers and a MGB probe were designed and synthesized according to the SPPV ORF068 gene in this study. The rapid detection method of SPPV TaqMan-MGB fluorescence quantitative PCR was established by optimizing the conditions. The results showed that this method could be used to detect SPPV-specific, but negative for sheep aphtha virus and porcine blister virus, and the detection limit was 10 copies / 渭 L, and the coefficient of variation of repeated test was less than 3. The above results show that the TaqMan-MGB qPCR method established in this study has the advantages of high specificity, high sensitivity, good repeatability, and can be used for rapid and accurate detection of SPPV.
【作者单位】： 中国农业科学院兰州兽医研究所/家畜疫病病原生物学国家重点实验室;深圳出入境检验检疫局动植物检验检疫技术中心;
【基金】：国家自然科学基金项目(31201892) 甘肃省国际科技合作专项(1504WKCA055、2016GS08219)
【分类号】：S852.654
本文编号：2186173
[Abstract]:In order to establish a rapid detection method for the (SPPV) gene of sheep pox virus, a pair of specific primers and a MGB probe were designed and synthesized according to the SPPV ORF068 gene in this study. The rapid detection method of SPPV TaqMan-MGB fluorescence quantitative PCR was established by optimizing the conditions. The results showed that this method could be used to detect SPPV-specific, but negative for sheep aphtha virus and porcine blister virus, and the detection limit was 10 copies / 渭 L, and the coefficient of variation of repeated test was less than 3. The above results show that the TaqMan-MGB qPCR method established in this study has the advantages of high specificity, high sensitivity, good repeatability, and can be used for rapid and accurate detection of SPPV.
【作者单位】： 中国农业科学院兰州兽医研究所/家畜疫病病原生物学国家重点实验室;深圳出入境检验检疫局动植物检验检疫技术中心;
【基金】：国家自然科学基金项目(31201892) 甘肃省国际科技合作专项(1504WKCA055、2016GS08219)
【分类号】：S852.654
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