乙脑病毒RT-PCR检测方法的建立及应用  

Development and application of RT-PCR assay of Japanese encephalitis virus

作　　者：高正琴[1] 岳秉飞[1] 贺争鸣[1] 

机构地区：[1]中国药品生物制品检定所,北京100050

出　　处：《药物分析杂志》2010年第10期1928-1931,共4页Chinese Journal of Pharmaceutical Analysis

基　　金：国家科技支撑计划(动物源性生物材料病毒安全性检测技术研究)(2008BAI54B06)

摘　　要：目的:建立乙脑病毒RT-PCR检测方法。方法:参照已发表的乙脑病毒全基因组序列,针对其衣壳蛋白C基因设计1对引物,提取乙脑病毒的RNA,逆转录为cDNA,进行PCR扩增,对检测方法进行优化,验证该方法的特异性及敏感性,并对90份临床样本进行检测。结果:所建立的乙脑病毒RT-PCR检测方法特异性和敏感性良好,以含乙脑病毒衣壳蛋白C基因的重组质粒DNA为模板,最低可检出10pgDNA。90份样本经RT-PCR检测,有23份扩增出特异性的目的条带。从小型猪脑组织中能分离鉴定出乙脑病毒。结论:已建立了乙脑病毒逆转录PCR检测方法,为乙脑病毒检测、流行病学调查奠定了基础。Objective：To establish an RT-PCR assay for detection of Japanese encephalitis virus（JEV）.Mehtods：A pairs of primers specific to capsid protein C gene of JEV SA14 were designed according to the published JEV complete genome sequence,with which the RNA of JEV was extracted and reversely transcribed to cDNA as a template for PCR amplification.Results：The established RT-PCR assay was optimized,verified for specificity and sensitivity,and its minimum limit using the recombination plasmid DNA containing JEV capsid protein C gene as template was 10 pg DNA.Ninety clinical samples were detected by the established RT-PCR assay,from thirty-three of which the specific target genes were amplified.JEV were isolated and identified from these brain tissues of mini-pigs.Conclusion：The established RT-PCR assay laid a foundation of detection and epidemiological investigation of JEV infection.

关 键 词：乙脑病毒 逆转录聚合酶链式反应 特异性 敏感性 检测 

分 类 号：R917[医药卫生—药物分析学]