荧光定量RT-PCR检测脑心肌炎病毒方法的建立及初步应用  

Development and preliminary application of fluorescent quantitative RT-PCR method for detection of encephalomyocarditis virus

作　　者：袁源[1,2] 许锬 郑朝共 容新宗 黄琰 苗松 张雪梅 

机构地区：[1]四川大学生命科学学院,成都610064 [2]成都蓉生药业有限责任公司,成都610041

出　　处：《四川大学学报（自然科学版）》2015年第1期205-209,共5页Journal of Sichuan University(Natural Science Edition)

基　　金：国家高技术研究发展计划(863计划)(2012AA021904)

摘　　要：根据GenBank中公布的脑心肌炎病毒(Encephalomyocarditis virus EMCV)3D基因保守区段设计并合成1对引物,以提取的EMCV核酸为模板,分别进行荧光定量RT-PCR扩增,优化反应体系及条件,建立脑心肌炎病毒荧光定量RT-PCR检测方法,并初步运用于静注人免疫球蛋白(IVIG)纳米膜过滤工艺去除EMCV效果的验证.结果显示该方法实验内和实验间的精密度均小于5%,最低检测限为102copies/μL;纳米膜过滤工艺可使样品中的EMCV滴度下降4Logs.Based on the conservative region of 3D gene of Encephalomyocarditis virus（EMCV ）published in GenBank,a pair of primers were Designed and synthesized.Use the extracted EMCV nucleic acid as a template for fluorescence quantitative RT-PCR amplification,optimization of reaction system and condi-tions.Establishencephalomyocarditis virus （EMCV）fluorescence quantitative RT-PCR detection meth-od,and applied to static note human immunoglobulin （IVIG）nano membrane filtration process to re-move EMCV effect validation.The results showed both of the precision of intra and inter assay was ＆lt;5% in Ct,and the lowest detective limit was 102 copies/μL.Nano membrane filtration process can make the samples of EMCV drops fell 4 logs.

关 键 词：荧光定量RT—PCR 脑心肌炎病毒 钠米膜 病毒去除 

分 类 号：Q78[生物学—分子生物学] R373[医药卫生—病原生物学]