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应用荧光RT-PCR检测亚欧型口蹄疫病毒 被引量：1

Detection of Euroasiatic Foot-and-Mouth Disease Virus by Real-time RT-PCR Assay
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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。 The universal probe and primers specific for FMDV Euroasiatic serotypes （including A, O, C and Asia-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 1.9×10^7copies 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.
作者 查成刚 吴绍强 林祥梅 韩雪清 刘建 贾广乐 梅琳
机构地区 南京农业大学动物医学学院 中国检验检疫科学研究院
出处 《中国动物检疫》 CAS 2009年第1期28-30,共3页 China Animal Health Inspection
基金 国家科技支撑计划(2006BAD06A14) 国家质检总局科研基金(2006IK037)资助
关键词 口蹄疫病毒 亚欧型 荧光RT-PCR 检测 Foot-and-Mouth Disease Euroasiatic serotype real-time PCR Detection.
分类号 S852.659.6 [农业科学—基础兽医学]
  • 相关文献

参考文献13

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共引文献104

同被引文献24

  • 1花群义,金宁一,周晓黎,董俊,杨云庆,徐自忠.应用实时荧光定量TaqManRT-PCR检测口蹄疫病毒[J].中国兽医学报,2005,25(2):116-119. 被引量：18
  • 2李永东,张常印,姜平,唐泰山,杨晓伟,杜以军.口蹄疫病毒实时RT-PCR检测方法的建立[J].中国人兽共患病学报,2006,22(3):212-215. 被引量：14
  • 3董志强,张志,李晓成,陈德坤,洪军,张燕霞,黄保续.口蹄疫检测技术研究进展[J].中国动物检疫,2007,24(3):48-50. 被引量：21
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引证文献1

二级引证文献7

中国动物检疫
2009年 第1期

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