蓝舌病病毒、流行性出血病病毒和帕利亚姆血清群病毒三重RT-qPCR检测方法的建立及应用
S852.65; [目的]建立蓝舌病病毒(Bluetongue virus,BTV)、流行性出血病病毒(Epizootic hemorrhagic disease virus,EHDV)及帕利亚姆血清群病毒(Palyam serogroup virus,PALV)快速筛查和诊断的三重RT-qPCR检测技术.[方法]选择BTV的NS3、EHDV的NS1以及PALV的VP7基因作为靶基因,设计3组特异性引物和探针,建立3种病毒的三重RT-qPCR检测方法.对检测方法的灵敏度、特异性和重复性进行验证,同时使用我国分离的BTV、EHDV与PALV不同血清型毒株和对应的阳性血液样本,与24个BTV血清型...
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| Published in | 华南农业大学学报 Vol. 42; no. 3; pp. 17 - 25 |
|---|---|
| Main Authors | , , , , , , , , |
| Format | Journal Article |
| Language | Chinese |
| Published |
云南省畜牧兽医科学院/云南省热带亚热带动物病毒病重点实验室,云南 昆明 650224
01.05.2021
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| Subjects | |
| Online Access | Get full text |
| ISSN | 1001-411X |
| DOI | 10.7671/j.issn.1001-411X.202007033 |
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| Abstract | S852.65; [目的]建立蓝舌病病毒(Bluetongue virus,BTV)、流行性出血病病毒(Epizootic hemorrhagic disease virus,EHDV)及帕利亚姆血清群病毒(Palyam serogroup virus,PALV)快速筛查和诊断的三重RT-qPCR检测技术.[方法]选择BTV的NS3、EHDV的NS1以及PALV的VP7基因作为靶基因,设计3组特异性引物和探针,建立3种病毒的三重RT-qPCR检测方法.对检测方法的灵敏度、特异性和重复性进行验证,同时使用我国分离的BTV、EHDV与PALV不同血清型毒株和对应的阳性血液样本,与24个BTV血清型及6个EHDV血清型参考毒株对该三重法的检测效果进行验证.[结果]三重RT-qPCR的扩增效率均可达90％以上,对3种病毒核酸拷贝数的检测下限均为10μL?1级别,与单重法的检测灵敏度相当;与阿卡斑病毒、小反刍兽疫病毒、口蹄疫病毒和牛流行热病毒之间无交叉反应;Ct值批内、批间变异系数均在2％以内;可检测出24种BTV血清型、6种EHDV血清型与3种PALV血清型,具有群特异性;可有效检测血液中的病毒核酸,检测结果与单重法一致.[结论]本研究建立的BTV、EHDV与PALV三重RT-qPCR具有良好的敏感性、特异性和重复性,可用于临床样本中对3种病毒的同时诊断与筛查. |
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| AbstractList | S852.65; [目的]建立蓝舌病病毒(Bluetongue virus,BTV)、流行性出血病病毒(Epizootic hemorrhagic disease virus,EHDV)及帕利亚姆血清群病毒(Palyam serogroup virus,PALV)快速筛查和诊断的三重RT-qPCR检测技术.[方法]选择BTV的NS3、EHDV的NS1以及PALV的VP7基因作为靶基因,设计3组特异性引物和探针,建立3种病毒的三重RT-qPCR检测方法.对检测方法的灵敏度、特异性和重复性进行验证,同时使用我国分离的BTV、EHDV与PALV不同血清型毒株和对应的阳性血液样本,与24个BTV血清型及6个EHDV血清型参考毒株对该三重法的检测效果进行验证.[结果]三重RT-qPCR的扩增效率均可达90％以上,对3种病毒核酸拷贝数的检测下限均为10μL?1级别,与单重法的检测灵敏度相当;与阿卡斑病毒、小反刍兽疫病毒、口蹄疫病毒和牛流行热病毒之间无交叉反应;Ct值批内、批间变异系数均在2％以内;可检测出24种BTV血清型、6种EHDV血清型与3种PALV血清型,具有群特异性;可有效检测血液中的病毒核酸,检测结果与单重法一致.[结论]本研究建立的BTV、EHDV与PALV三重RT-qPCR具有良好的敏感性、特异性和重复性,可用于临床样本中对3种病毒的同时诊断与筛查. |
| Author | 肖雷 廖德芳 李占鸿 谢佳芮 朱建波 杨恒 李华春 杨振兴 李卓然 |
| AuthorAffiliation | 云南省畜牧兽医科学院/云南省热带亚热带动物病毒病重点实验室,云南 昆明 650224 |
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| Author_FL | LIAO Defang YANG Zhenxing LI Zhanhong XIAO Lei XIE Jiarui |
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| DocumentTitle_FL | Establishment and application of triplex RT-qPCR for detection of bluetongue virus, epizootic hemorrhagic disease virus and Palyam serogroup virus |
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| Keywords | 帕利亚姆血清群病毒 病毒检测 蓝舌病病毒 三重RT-qPCR 流行性出血病病毒 |
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| Publisher | 云南省畜牧兽医科学院/云南省热带亚热带动物病毒病重点实验室,云南 昆明 650224 |
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| Title | 蓝舌病病毒、流行性出血病病毒和帕利亚姆血清群病毒三重RT-qPCR检测方法的建立及应用 |
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