黄热病毒实时荧光定量PCR方法的建立  

Establishment on TaqMan real-time fluorescent quantitative PCR for detecting yellow fever virus

作　　者：祁倩[1] 劳小荣[1] 陈蔼珍[1] 熊建英[1] 周旋[1] 张其威[1] 朱利[1] 李凌[1] 万成松[1] 赵卫[1] 

机构地区：[1]南方医科大学公共卫生与热带医学院中心实验室,广东广州510515

出　　处：《现代预防医学》2013年第15期2863-2866,2871,共5页Modern Preventive Medicine

基　　金：广东省社会发展领域科技计划项目(2010B031000005;2011B031500015);广州市科技亚运专项(2010U1-E00591)

摘　　要：目的建立黄热病毒的实时荧光定量PCR检测技术,以实现黄热病毒感染早期的筛选和检测。方法以带有黄热病毒基因质粒为阳性模板,采用TaqMan探针法,制作标准曲线,以实现对黄热病毒的简单快速的高灵敏度、高特异性实时检测。结果该方法检测的最低拷贝数可以达到3.457copies/μl。组内及组间变异系数均低于5%,证明该方法具有良好的敏感性及稳定性。以1～4型登革病毒和乙脑病毒基因组为模板,检测结果为阴性,证明该方法有良好的特异性。结论该方法可快速、灵敏、特异、定量检测黄热病毒,能用于黄热病毒感染的早期诊断。OBJECTIVE To establish a specific, sensitive method with real-time fluorescence quantitative PCR assay for the rapid detection of yellow fever virus （YFV）. METHODS In order to test yellow fever virus for simple and fast real-time detection with high sensitivity and high specificity, we selected the gene plasmid of yellow fever virus as the positive template. Used TaqMan probe method, we produced standard curve. RESULTS The sensitivity of this way was 3.457 copies / μl.In the group and variation coefficients between groups were less than 5%. The results showed that the method had a good sensitivity and stability. According to the mix of experiment with dengue virus type 1 to 4 and Japanese encephalitis virus , we proved this method had a good specificity. CONCLUSION The TaqMan real-time fluorescent quantitative PCR assay is a rapid, sensitive and specific method for the quantitative detection of YFV; it’s applicable for the early clinical diagnosis of YFV and there are very good social and economic benefits.

关 键 词：黄热病毒 实时荧光定量PCR 检测 

分 类 号：R373.3[医药卫生—病原生物学]