人免疫缺陷病毒Ⅰ型gag、env、rev mRNA检测  

Detection of human immunodeficiency virus type Ⅰ gag、env、rev mRNA

作　　者：姜拥军[1] 尚红[1] 康辉[1] Gregory F.Burton 王亚男[1] 

机构地区：[1]中国医科大学第一临床学院艾滋病研究室,沈阳110001 [2]UT Brigham Young University

出　　处：《中华检验医学杂志》2002年第5期296-298,共3页Chinese Journal of Laboratory Medicine

摘　　要：目的　动态检测人免疫缺陷病毒Ⅰ型 (HIV 1)基因gag、env、revmRNA水平。方法　将插入有HIV 1竞争模板的pSPI质粒DNA转化入感受态菌STBL 2 ,进行扩增。以T7RNA聚合酶体外转录出HIV 1mRNA竞争模板。用 3对引物对HIV 1RNA标本进行定量竞争聚合酶链反应 (QC RT PCR) ,检测HIV 1gag、env、revmRNA水平。结果　HIV 1ⅢB感染的标本中 ,HIV 1gag、env和revmRNA的水平分别为 36 0 0 0拷贝 / μl,980 0拷贝 / μl和 910 0拷贝 / μl。此方法可动态定量检测HIV 1gag、env、revmRNA水平。结论　该方法简便易行 ,费用低廉 ,适用于实验室HIV致病机理、药物筛选和病毒复制状况的研究。Objective To dynamically detect the level of human immunodeficiency virus type I gag?env?rev mRNA. Methods pSPI plasmid DNA was transformed into competent cell STBL 2 and amplified. Linear pSPI plasmid DNA was used to produce competitive RNA template by in vitro transcription with T 7 RNA polymerase. The level of HIV 1 gag?env and rev mRNA were determined separately by QC RT PCR with three pairs of primers. Results The levels of HIV 1 gag?env and rev mRNA were 36 000 copies/μl, 9 800 copies/μl and 9 100 copies /μl respectively. The method could be used to dynamically quantify HIV 1 gag?env?rev mRNA. Conclusions This method is simple, easy and cheaper. Quantitative detection of HIV 1 gag?env?rev mRNA can be applied to study HIV pathogenesis, anti HIV drug screening, and HIV replication status.

关 键 词：人免疫缺陷病毒-Ⅰ 病毒基因 信使核糖核酸 艾滋病 AIDS 

分 类 号：R446.5[医药卫生—诊断学]