结合RT-PCR体外扩增检测HCVssRNA的研究  

The Study of Amplifing HCV ssRNA by RT-PCR Combined With Transcription in vitro

作　　者：胡俊[1] 董长垣[1] 汪凡军[1] 陈晓[1] 郭淑芳[1] 张蔚英[1] 

机构地区：[1]武汉大学医学院病毒研究所,湖北武汉430071

出　　处：《中国病毒学》2004年第3期214-216,共3页Virologica Sinica

基　　金：湖北省科技厅重点项目(12410135)

摘　　要：本研究旨在以HCV为平台,在简化RT-PCR基础上,结合体外转录,建立一种特异、高效、简便的检测血清中HCVRNA的体外转录合成系统。本法扩增终产物为特定极性的ssRNA,其特异性经凝胶电泳和斑点杂交确定;RNA定量分析结果显示本法核酸扩增效率高于巢式PCR近20倍。The purpose of this study is to establish a new method which HCV RNA can be used to detect sensitively and specifically through efficient single strand RNAs(ssRNAs) amplification. The sera from the chronic Hepatitis C patients were used as specimens. Single step reverse transcription-PCR (SRT-PCR) was performed prospectively, with a designed set of primers which defined the sequence unique to the 5' noncoding region of the HCV genome, a great deal of specific ssRNAs were synthesized in the presence of T7RNA polymerase and other appropriate conditions within five hours. Comparing with the nested-PCR, the quantity of the products in our method was nearly 20-fold higher and its specificity was verified by the electrophoresis on agarose gel and the dot hybridization with the probe.

关 键 词：丙型肝炎病毒 逆转录PCR 体外转录 

分 类 号：R446.5[医药卫生—诊断学]