Sevall J S, Prince H, Garratty G, O'Brien W A, Zack J A
Specialty Laboratories, Inc., Santa Monica, CA 90404.
Clin Chem. 1993 Mar;39(3):433-9.
A clinical procedure for rapid detection of human immunodeficiency virus type 1 (HIV-1) by DNA amplification is demonstrated. The rapid procedure reduces handling requirements and amplification time and eliminates use of radioactivity for the detection of the amplification product. Total leukocyte lysates are the amplification substrates. Two conserved regions in the HIV-1 genome are amplified by 45 cycles of a two-temperature thermal cycle and the amplification products are detected by ultraviolet light after electrophoresis on agarose gels. Twenty-four specimens clinically diagnosed by detection of antibody (IgG) to HIV-1 were confirmed by the rapid DNA amplification procedure. In a blind study, 56 samples positive for HIV-1 DNA were detected in 503 individuals by the current classical polymerase chain reaction method; the same 56 positive samples were also detected by the rapid amplification protocol. No false-positive or false-negative results were obtained. The turnaround time for analysis has been reduced to < 24 h without compromising test results.
本文展示了一种通过DNA扩增快速检测1型人类免疫缺陷病毒(HIV-1)的临床方法。该快速方法减少了操作要求和扩增时间,并消除了用于检测扩增产物的放射性物质的使用。全白细胞裂解物作为扩增底物。通过两温度热循环45个周期扩增HIV-1基因组中的两个保守区域,并在琼脂糖凝胶上电泳后通过紫外光检测扩增产物。通过快速DNA扩增程序确认了24个经检测HIV-1抗体(IgG)临床诊断的标本。在一项盲法研究中,通过当前经典聚合酶链反应方法在503名个体中检测到56个HIV-1 DNA阳性样本;快速扩增方案也检测到了相同的56个阳性样本。未获得假阳性或假阴性结果。分析的周转时间已缩短至<24小时,且不影响检测结果。