| 应用荧光RT-PCR检测亚欧型口蹄疫病毒 |
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| 引用本文： | 查成刚,吴绍强,林样梅,韩雪清,刘建,贾广乐,梅琳.应用荧光RT-PCR检测亚欧型口蹄疫病毒[J].中国动物检疫,2009,26(1):28-30. |
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| 作者姓名： | 查成刚 吴绍强 林样梅 韩雪清 刘建 贾广乐 梅琳 |
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| 作者单位： | 1. 南京农业大学动物医学学院,江苏,南京,210095;中国检验检疫科学研究院,北京,朝阳,100029
2. 中国检验检疫科学研究院,北京,朝阳,100029 |
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| 基金项目： | 国家科技支撑计划，国家质检总局科研项目 |
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| 摘 要： | 根据口蹄疫病毒基因组的5’非翻译区序列比较保守的特点,采用软件设计亚欧型(A、O、C及Asia-1型)FMDV通用的引物和探针,经对反应条件和反应体系进行优化,建立了亚欧型FMDV的实时荧光RT-PCR检测方法。研究表明,该方法检测阳性质粒模板的线性范围为1.9×107-1.9×10拷贝,最低可检测19个拷贝。通过对FMDV各血清型(A、O、C、Asia-1及SAT-1,2,3型)的检测,证实该方法对亚欧型FMDV具有良好的特异性,能有效区分亚欧型和南非型FMDV感染。本研究为亚欧型口蹄疫的快速诊断和流行病学调查提供了新的方法。
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| 关 键 词： | 口蹄疫病毒 亚欧型 荧光RT-PCR 检测 |
Detection of Euroasiatic Foot-and-Mouth Disease Virus by Real-time RT-PCR Assay |
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| Zha Chenggang,Wu Shaoqiang,Lin Xiangmei,Han Xueqing,Liu Jian,Jia Guangle,Mei Lin.Detection of Euroasiatic Foot-and-Mouth Disease Virus by Real-time RT-PCR Assay[J].China Journal Of Animal Quarantine,2009,26(1):28-30. |
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| Authors: | Zha Chenggang Wu Shaoqiang Lin Xiangmei Han Xueqing Liu Jian Jia Guangle Mei Lin |
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| Institution: | 1.College of Veterinary Science,Nanjing Agricultural University,Nanjing 210095;2.Chinese Academy of Inspection and Quarantine,Beijing 100029) |
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| Abstract: | The universal probe and primers specific for FMDV Euroasiatic serotypes(including A,O,C and Asi-a-1) were designed and synthesized according to the 5’ untranslated region(UTR) of FMDV genomic sequence available on GenBank.The reaction conditions were optimized for sensitivity and specificity.Results indicated that,the dynamic range of the developed FMDV real-time PCR assay is from 19 copies to 1.9×107copies of the constructed plasmids inserted with the targeted FMDV DNA fragment,and the sensitivity is as low as 19 copies of plasmid DNA.The specificity of real-time RT-PCR method was verified when using the total RNA of FMDV South Africa serotypes as template and no amplification plot were found.The developed method could be used for rapid diagnosis and epidemiological survey of Euroasiatic FMD in the field. |
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| Keywords: | Foot-and-Mouth Disease Euroasiatic serotype real-time PCR Detection |
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