Ou C Y, Kwok S, Mitchell S W, Mack D H, Sninsky J J, Krebs J W, Feorino P, Warfield D, Schochetman G
Center for Infectious Diseases, Centers for Disease Control, Atlanta, GA 30333.
Science. 1988 Jan 15;239(4837):295-7. doi: 10.1126/science.3336784.
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.
通过一种称为聚合酶链反应的选择性DNA扩增技术,在从血清反应阳性者外周血单个核细胞(PBMC)中分离出的DNA中直接鉴定出了人类免疫缺陷病毒(HIV-1)的前病毒序列,但在从该病毒血清反应阴性者PBMC中分离出的DNA中未检测到。使用来自HIV-1基因组多个区域的引物对,以实现前病毒检测的最大灵敏度。在通过共培养分离出病毒的血清反应阳性同性恋男性的100%DNA样本中检测到了HIV-1序列,但在血清反应阴性、病毒培养阴性的对照组人员的DNA样本中均未检测到。然而,在血清反应阳性、病毒培养阴性的同性恋男性的64%DNA样本中检测到了HIV-1序列。这种DNA扩增方法能够在3天内获得结果,而病毒分离则需要长达3至4周。因此,该方法可作为确定HIV-1感染的常规手段,用于补充或替代病毒分离。