逆转录—聚合酶链反应在HIV-1感染诊断中的应用

Using RT-PCR Method to Diagnose HIV-1 infection

[目的 ]评价逆转录 -聚合酶链反应 (RT PCR)在HIV 1感染诊断中的应用。[方法 ]采用RNA一步法试剂盒提取RNA。用 3套分别来自于LTR gag、pol、env不同区域的序列引物进行嵌套式PCR扩增。[结果 ]对 2 4例血清标本进行检测 ,其中 15例为HIV感染者 ;1例为免疫印迹试验可疑待确定 ,但经过 1年的随访可排除 ;另外 8例为正常者。经过巢式扩增 ,15份阳性标本中有 14份扩增出两条或两条以上核酸片段 ,1份未扩增出任何一条片段 ,其敏感性达93 3 % ( 14 /15 )。其中 6份肝素抗凝标本env区域均未扩增出片段。阴性标本无片段 ,其特异性为 10 0 % ( 8/8)。[结论 ]RT PCR具有高敏感性和特异性 ,可用于窗口期HIV感染的诊断及HIV 1阳性母亲所生婴儿的HIV检测 ,可作为免疫印迹试验的补充和辅助诊断。

Objective\] To evaluate the application of Nested RT PCR in detection of HIV 1 infection.\ RNA was extracted following the instruction of RNA one step kit. Nested PCR was performed by using three HIV 1 primer sets which amplified a fragment in the LTR gag,pol and env gene respectively. \ 24 samples including 15 HIV 1 seropositives, one indeterminated sample, and 8 seronegatives were tested by nested RT PCR. 14 seropositive samples were amplified two DNA fragments or more. Only 1 sample show negative as no DNA fragment was seen. According the testing algorithm, the sensitivity and specificity of the test was 93.3% and 100% respectively. \[Conclusion\] Nested RT PCR is high sensitivity and specificity that can be used to diagnose of HIV infection especially in window period and the child born by HIV 1 seropositive mother.