摘要
根据HIV-1 gag.pol和env基因序列设计了套式聚合酶链反应(nesled polymerasechain reaction,nPCR)引物。将外周血单个核细胞裂解液先用外侧引物扩增,其产物再经内侧引物扩增,直接走凝胶电泳判定结果。nPCR的敏感性。较常规PCR高100～1000倍,可检测到0.5fg质粒DNA和50μ1HIV-1感染者外周血样中的HIV基因。用本法检测63名HIV-1感染者全为阳性,而61名健康人和可疑者均为阴性,后者经追踪检测抗体排除了HIV-1感染。实验证明,nPCR是一种敏感性极高、特异性很强、操作简便易行的HIV-1基因检测技术。它已经在早期诊断和外周血病毒含量检测中发挥了重要作用,而且有着更广阔的应用前景。
Four sets of nested primers corresponding to HIV-1 gag, pol and env genes were designed and synthesized. Peripheral blood mononuclear cell lysates were first amplified with outer primers. Its products were fur-ther amplified with the inner nested primers. The final amplified band were detected by agarose gel electrophoresis. The nested PC R can detect 0.5 fg of plasmid DNA or cell lysates from 50μl of blood, it's sensiti-vity is 100 to l ,000 times higher than the standard PCR. All 63 HIV-1 infected patients were found to be positive. 55 HIV-l seronegative and 6 indeterminate individuals were found negative, the latter group were all diagnosed as HIV-1 seronegative after 3 to 6 month follow up. Our data proved that nested PCR is a sensitive, specific and simple method to detect HIV DNA in clinical samples, which can be used for the early diagnosis and early confirmation of HIV infection.
出处
《病毒学报》
CAS
CSCD
北大核心
1993年第3期261-268,共8页
Chinese Journal of Virology
关键词
套式
聚合酶链反应
艾滋病毒
诊断
Nested PCR, HIV-1 gene detection, Peripheral blood mo-nonuclear cells.
