| 小反刍兽疫实时荧光PCR检测方法的建立及应用 |
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| 引用本文： | 吴绍强,林祥梅,刘忠清,王彩霞,韩雪清,刘建,贾广乐,梅琳,王建峰,查成刚.小反刍兽疫实时荧光PCR检测方法的建立及应用[J].黑龙江畜牧兽医,2009(5). |
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| 作者姓名： | 吴绍强 林祥梅 刘忠清 王彩霞 韩雪清 刘建 贾广乐 梅琳 王建峰 查成刚 |
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| 作者单位： | 吴绍强,林祥梅,王彩霞,韩雪清,刘建,贾广乐,梅琳,王建峰,查成刚(中国检验检疫科学研究院,动植物检疫研究所,北京100029);刘忠清(西藏出入境检验检疫局,西藏,拉萨,850000)
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| 摘 要： | 以小反刍兽疫病毒N基因序列为靶基因,通过设计引物及TaqMan探针建立快速检测小反刍兽疫的荧光PCR方法.在线BLAST分析结果表明:所设计的引物特异性强,可区分小反刍兽疫病毒及牛瘟病毒感染;所建立的方法检测线性范围为1～1×106拷贝质粒DNA,灵敏度可达10拷贝质粒DNA,是常规PCR法的100倍.应用所建立的方法对32份采自西藏疫区的组织样品进行检测,说明疫情在西藏没有扩散.
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| 关 键 词： | 小反刍兽疫病毒 荧光PCR 检测 |
Establishment and application of real time PCR method for detection of peste des petits ruminants |
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| WU Shao-qiang,LIN Xiang-mei,LIU Zhong-qing,et al.Establishment and application of real time PCR method for detection of peste des petits ruminants[J].Heilongjiang Animal Science And veterinary Medicine,2009(5). |
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| Authors: | WU Shao-qiang LIN Xiang-mei LIU Zhong-qing |
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| Institution: | 1.Chinese Academy of Inspection and Quarantine;Beijing 100029;China;2.Tibet Entry-exit Inspection and Quarantine;Lasa 850000;China |
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| Abstract: | The real time PCR method was developed to detect peste des petits ruminants(PPR) contamination in animals and animal products.The primers and TaqMan probe were designed and selected to amplify the species-specific N gene of PPRV.The sensitivity and specificity of the developed method were examined using the plasmid containing the targeted N gene fragment as template.The online BLAST analysis showed that the primers were specific enough to distinguish PPRV from rinderpest virus(RPV) infection.The dynamic ran... |
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