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

作　　者：刘阳[1,2] 史子学[1] 马玉堃[2] 王皓婷[1] 王宗尧[1] 邵东华[1] 魏建超[1] 王少辉[1] 李蓓蓓[1] 王水明[3] 刘学辉[3] 马志永[1] 

机构地区：[1]中国农业科学院上海兽医研究所兽医公共卫生研究室,上海200241 [2]黑龙江大学生命科学学院,哈尔滨150080 [3]南京出入境检验检疫局,南京211136

出　　处：《病毒学报》2012年第5期567-571,共5页Chinese Journal of Virology

基　　金：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)

摘　　要：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102～1010,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。In order to establish a rapid and accurate method for the detection of Ebola virus （EBOV）, the primers used in SYBR Green Ⅰ real-time RT-PCR were designed based on the EBOV NP gene sequences published in GenBank. The SYBR Green Ⅰ real-time RT-PCR was established and optimized for the detection of EBOV. The EBOV RNA that was transcribed in vitro was used as a template. The sensitivity of this method was found to reach 1.0 × 10^2 copies/μL and the detection range was 10^2 × 10^10. No cross reaction with RNA samples from Marburg virus, Dengue virus, Xinjiang hemorrhagic fever virus, Japanese encephalitis virus, Influenza virus （H1N1 and H3N2） and Porcine reproductive and respiratory syndrome virus E genomic RNA was found.. The method would be useful for the detection and monitoring of EBOV in China.

关 键 词：埃博拉病毒 荧光定量RT-PCR SYBR Green Ⅰ 检测 

分 类 号：S183.9[农业科学—农业基础科学]