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(四川大学生命科学学院)；许锬(成都蓉生药业有限责任公司)；郑朝共(成都蓉生药业有限责任公司)；容新宗(成都蓉生药业有限责任公司)；黄琰(成都蓉生药业有限责任公司)；苗松(成都蓉生药业有限责任公司)；张雪梅(成都蓉生药业有限责任公司)
Author：YUAN Yuan(College of Life Sciences, Sichuan University)；XU Tang(Chengdu Ronsen Pharmaceuticals Co., Ltd.)；ZHENG Chao-Gong(Chengdu Ronsen Pharmaceuticals Co., Ltd.)；RONG Xin-Zong(Chengdu Ronsen Pharmaceuticals Co., Ltd.)；HUANG Yan(Chengdu Ronsen Pharmaceuticals Co., Ltd.)；MIAO Song(Chengdu Ronsen Pharmaceuticals Co., Ltd.)；ZHANG Xue-Mei(Chengdu Ronsen Pharmaceuticals Co., Ltd.)
收稿日期：2014-02-20 年卷（期）页码：2015,52(1):205-209
期刊名称：四川大学学报: 自然科学版
Journal Name：Journal of Sichuan University (Natural Science Edition)
关键字：荧光定量RT PCR; 脑心肌炎病毒; 钠米膜; 病毒去除
Key words：Quantitative RT PCR; Encephalomyocarditis virus; nano membrane; Virus removal
基金项目：国家高技术研究发展计划(863计划)(2012AA021904)
中文摘要
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102 copies/ μL; 纳米膜过滤工艺可使样品中的EMCV滴度下降4Logs.
英文摘要
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 conditions. Establishencephalomyocarditis virus (EMCV) fluorescence quantitative RT PCR detection method, and applied to static note human immunoglobulin (IVIG) nano membrane filtration process to remove EMCV effect validation. The results showed both of the precision of intra and inter assay was ＜5% in Ct, and the lowest detective limit was 102copies/μL. Nano membrane filtration process can make the samples of EMCV drops fell 4 logs.
上一条：菠菜43 kD叶绿素a结合蛋白编码基因克隆及原核表达条件优化
【关闭】
