HIV—Pol基因的套式PCR检测

DETECTION OF HIV-1 POL GENE BY NESTED POLYMERASE CHAIN REACTION

根据ＨＩＶ—１ ＭＮ株Ｐｏｌ基因序列设计了套式聚合酶链反应（Ｎｅｓｔｅｄｐｏｌｙｍｅｒａｓｅｃｈａｉｎｒｅａｃｔｉｏｎ，ｎＰＣＲ）引物，将外周血单核细胞ＤＮＡ粗提物先以一对外侧引物扩增，其产物再以一对内侧引物扩增，两次扩增后的产物直接经电泳判定结果。ｎＰＣＲ在保持常规ＰＣＲ特异性的同时，其敏感性较单对引物ＰＣＲ高１００倍以上，可检出约１０个分子的目的基因片段。探讨了ＨＩＶ—１ Ｐｏｌ基因套式ＰＣＲ检测在ＨＩＶ—１感染的早期诊断、感染确认及病毒分离中的应用，表明本方法是一种敏感性及特异性俱佳，结果判断简单的基因检测技术，有应用前景。

group of nested primers corresponding to HIV-1 MN strain Pol gene was designed and synthesized .Peripheral blood mononuclear cell lysates were first amplified with outer primers.Its products further amplified with the inner nested primers.The final amplified products were detected by agarose gel electrophoresis. The sensitivity of nested PCR is 100 to 1000 times higher than the standard PCR and can detect 10 molecules of template DNA.Our data showed that the HIV-1 Pol gene detection by nested PCR is valuable in early diagnosis ,confirmation of HIV 1 infecton and HIV-1 isolation.It is a simple gene detecting method with high specificity and sensitivity.