| 狂犬病病毒快速荧光RT-PCR检测方法研究与应用 |
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| 引用本文： | 高志强,张鹤晓,柏亚铎,赖平安,谷强,凌凤俊,张伟,张向东,段生涛.狂犬病病毒快速荧光RT-PCR检测方法研究与应用[J].中国兽医杂志,2010,46(1). |
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| 作者姓名： | 高志强 张鹤晓 柏亚铎 赖平安 谷强 凌凤俊 张伟 张向东 段生涛 |
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| 作者单位： | 北京出入境检验检疫局,北京,通州,101113 |
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| 基金项目： | 北京市科技计划:国家科技计划项目衔接-科技奥运专项，国家质量监督检验检疫总局科技计划 |
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| 摘 要： | 采用TaqMan方法,根据古典狂犬病病毒非编码区及N基因编码区序列,设计合成多对引物和多条探针,通过对引物、探针的筛选,反应条件的选择和优化,建立了古典狂犬病病毒荧光RT-PCR检测技术。与病毒分离鉴定、常规RT-PCR试验比较表明,建立的荧光PCR检测技术快速、敏感,检测时限3 h以内。特异性试验表明本方法与犬瘟热病毒,犬细小病毒,犬Ⅰ型腺病毒不发生交叉反应。初步动物感染试验及定量检测研究及表明,本方法可用于感染动物不同组织样品中病毒的相对定量。对132份临床样品和4种商品化疫苗进行检测研究表明,本方法适用于样品中狂犬病病毒的直接检测以及疫苗中活病毒或灭活病毒的检测。
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| 关 键 词： | 狂犬病病毒 荧光RT-PCR 研究与应用 |
Studies and application of the fast detection rabies virus (RV) by real time RT-PCR |
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| GAO Zhi-qiang,ZHANG He-xiao,BAI Ya-duo,LAI Ping-an,GU Qiang,LING Feng-jun,ZHANG Wei,ZHANG Xiang-dong,DUAN Sheng-tao.Studies and application of the fast detection rabies virus (RV) by real time RT-PCR[J].Chinese Journal of Veterinary Medicine,2010,46(1). |
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| Authors: | GAO Zhi-qiang ZHANG He-xiao BAI Ya-duo LAI Ping-an GU Qiang LING Feng-jun ZHANG Wei ZHANG Xiang-dong DUAN Sheng-tao |
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| Institution: | GAO Zhi-qiang,ZHANG He-xiao,BAI Ya-duo,LAI Ping-an,GU Qiang,LING Feng-jun,ZHANG Wei,ZHANG Xiang-dong,DUAN Sheng-tao(Beijing Entry-exit Inspection , Quarantine Bureau,Beijing 101113,China) |
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| Abstract: | In this study,a real-time RT-PCR(RRT-PCR) was developed to detect classical rabies virus(RV) using Taq Man technique by the selection and optimization of primers,probes and reaction conditions.Compared with virus isolation(VI) and normal RT-PCR,the developed RRT-PCR was fast and sensitive,with the detection time less than 3 hours.The results also clearly demonstrated high specificity of developed RRT-PCR for the detection of RV by testing canine distemper virus,canine parvovirus and canine adenovirus type 1... |
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| Keywords: | rabies virus real-time RT-PCR studies and applicance |
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