DNA AMPLIFICATION FOR DIRECT DETECTION OF HIV-1 IN DNA OF PERIPHERAL-BLOOD MONONUCLEAR-CELLS
DNA AMPLIFICATION FOR DIRECT DETECTION OF HIV-1 IN DNA OF PERIPHERAL-BLOOD MONONUCLEAR-CELLS
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DOI：
10.1126/science.3336784
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发表时间：
1988-01-15
期刊：
影响因子：
56.9
通讯作者：
SCHOCHETMAN, G
中科院分区：
文献类型：
--
作者：
OU, CY;KWOK, S;SCHOCHETMAN, G
通过一种选择性的DNA扩增技术，称为聚合酶链反应，人免疫缺陷病毒（HIV-1）的前病毒序列被确定直接从外周血单核细胞（PBMC）的人血清阳性，但不是在DNA分离的人PBMCs的血清阴性的病毒。来自HIV-1基因组多个区域的引物对用于实现前病毒检测的最大灵敏度。HIV-1序列检测100%的DNA标本从血清阳性，同性恋男子的病毒是通过共培养分离，但在没有从一个对照组的血清阴性，病毒培养阴性的人的DNA标本。然而，HIV-1序列中检测到64%的DNA标本血清阳性，病毒培养阴性的同性恋男子。这种DNA扩增方法可以在3天内获得结果，而病毒分离需要长达3至4周。因此，该方法可用于补充或取代病毒分离作为确定HIV-1感染的常规手段。
By means of a selective DNA amplification technique called polymerase chain reaction, proviral sequences of the human immunodeficiency virus (HIV-1) were identified directly in DNA isolated from peripheral blood mononuclear cells (PBMCs) of persons seropositive but not in DNA isolated from PBMCs of persons seronegative for the virus. Primer pairs from multiple regions of the HIV-1 genome were used to achieve maximum sensitivity of provirus detection. HIV-1 sequences were detected in 100% of DNA specimens from seropositive, homosexual men from whom the virus was isolated by coculture, but in none of the DNA specimens from a control group of seronegative, virus culture-negative persons. However, HIV-1 sequences were detected in 64% of DNA specimens from seropositive, virus culture-negative homosexual men. This method of DNA amplification made it possible to obtain results within 3 days, whereas virus isolation takes up to 3 to 4 weeks. The method may therefore be used to complement or replace virus isolation as a routine means of determining HIV-1 infection.