SYBR Green Ⅰ荧光定量RT-PCR检测埃博拉病毒方法的建立Development of SYBR Green Ⅰ real-time RT-PCR for the Detection of Ebola Virus
刘阳;史子学;马玉堃;王皓婷;王宗尧;邵东华;魏建超;王少辉;李蓓蓓;王水明;刘学辉;马志永;
摘要(Abstract)：
为了建立一种快速准确的检测埃博拉病毒(EBOV)亚型的方法。本研究根据GenBank中公布的EBOV NP基因序列,通过设计引物和优化反应条件,建立了一种SYBR GreenⅠ荧光定量RT-PCR检测方法检测EBOV。以体外转录的EBOV RNA为模板进行试验,该方法检测的灵敏度可以达到1.0×102个拷贝/μL,检测范围达到9个数量级为102个拷贝/μL,检测范围达到9个数量级为102～102～10(10),可检测5种亚型EBOV。建立的方法对马尔堡病毒(MARV)、登革病毒(DENV)、新疆出血热病毒(XHFV)、乙型脑炎病毒(JEV)、流感病毒(H1N1和H3N2)和猪繁殖和呼吸综合征病毒(PRRSV)E基因组RNA无非特异性扩增。本文将荧光定量RT-PCR技术用于埃博拉病毒的定量检测中,并且建立了EBOV SYBRGreenⅠ荧光定量RT-PCR检测方法。
关键词(KeyWords)： 埃博拉病毒;荧光定量RT-PCR;SYBR Green Ⅰ;检测
基金项目(Foundation): 江苏出入境检验检疫局科研项目(2011KJ03);; 公益性行业(农业)科研专项经费(200903037-06)
作者(Authors): 刘阳;史子学;马玉堃;王皓婷;王宗尧;邵东华;魏建超;王少辉;李蓓蓓;王水明;刘学辉;马志永;
DOI: 10.13242/j.cnki.bingduxuebao.002323
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