滤纸片干血斑HIV-1 DNA检测方法的建立  

Establishment of a method to detect human immunodeficiency virus type 1 DNA on dried blood spots

作　　者：李秋荣[1] 邢文革[2] 姚均[2] 龙小山[2] 潘品良[2] 张麒[2] 蒋岩[2] 

机构地区：[1]北京朝阳区疾病预防控制中心,北京100021 [2]中国疾病预防控制中心性病艾滋病预防控制中心,北京100050

出　　处：《中国艾滋病性病》2007年第6期521-524,共4页Chinese Journal of Aids & STD

摘　　要：目的探索用滤纸片作血液样本的载体,建立一种简单、经济、敏感的滤纸片干血斑艾滋病病毒Ⅰ型(HIV-1)DNA检测方法。解决HIV-1感染者全血样本采集、运输和储存过程中存在的实际困难。方法首先用滤纸片FTA卡收集血液制成滤纸片干血斑,处理提取DNA后,针对HIV的env、gag、pol区,设计三对高度保守引物,套式PCR扩增HIV-1 DNA。进一步用HIV-1质粒DNA检测该方法的最低检测限。结果摸索出扩增滤纸片干血斑HIV-1 DNA的最佳扩增条件;滤纸片干血斑样本pol区最低检测限是6拷贝/μl,env区8拷贝/μl,gag区10拷贝/μl。结论可以用滤纸片FTA卡收集HIV-1感染者的血液,制成滤纸片干血斑后检测HIV-1 DNA。该方法操作简便、经济、敏感,建议作为HIV-1抗体检测的辅助诊断方法。Objective To establish a simple, cost effective and sensitive method to detect human immunodeficiency virus type 1 （HIV-1）DNA on dried blood spots（DBSs）by collecting blood on filter paper. Methods Blood was collected on FTA cards to get dried blood spots. Using three primer sets, the disc of DBSs was amplified by Nested PCR. HIV-1 plasmid DNA was used to determine the low detection limit of the assay. Results Optimization of PCR system and procedure of testing HIV-1 DNA on DBSs was conducted.The lowest limit of HIV-1 DNA pol detection with filter paper was 6 copies/μl,env region was 8 copies/μl,and gag region was 10 copies/μl.Conclusions It is feasible to collect blood of HIV- 1 positive individuals on FTA. The method of detecting HIV-1 DNA on dried blood spots is simple, cost effective and sensitive, So it is proposed that this method be used as an auxiliary diagnostic tool of HIV-1 antibody test.

关 键 词：滤纸片干血斑 艾滋病病毒Ⅰ型DNA套式聚合酶链反应 

分 类 号：R373.9[医药卫生—病原生物学]