Conway B, Bechtel L J, Adler K A, D'Aquila R T, Kaplan J C, Hirsch M S
Department of Microbiology & Immunology, University of Ottawa, Canada.
Mol Cell Probes. 1992 Jun;6(3):245-9. doi: 10.1016/0890-8508(92)90023-q.
We have compared spot-blot methodology with a recently developed rapid microtitre plate assay for the specific detection of HIV-1 PCR products. We have studied blood specimens isolated from HIV-1 infected individuals (48 asymptomatic and 56 symptomatic patients). Mononuclear cells were isolated, lysed and processed for PCR. Both PCR product detection methods were carried out in parallel on all amplified samples. HIV-1 sequences were detected by spot-blot or microtitre plate hybridization in samples taken from 42/48 asymptomatic and 53/56 symptomatic subjects. Concordant results between the two detection methods were observed for 90 samples, with 81 positive and nine negative assays. On repeat evaluation of the 14 discordant samples, nine showed concordant positive results, near the limit of detection of the assay. Serial dilutions of ACH-2 cells were amplified, and the PCR products were detected using the microtitre plate assay, yielding semi-quantitative results. The sensitivity of this simple, rapid assay compares with that of more laborious DNA detection systems. This may become a useful tool in HIV-1 research and in the clinical care of seropositive individuals.
我们已将斑点印迹法与最近开发的用于特异性检测HIV-1 PCR产物的快速微量滴定板检测法进行了比较。我们研究了从HIV-1感染个体（48例无症状患者和56例有症状患者）分离出的血液标本。分离单核细胞，进行裂解并处理以用于PCR。两种PCR产物检测方法在所有扩增样本上并行进行。通过斑点印迹或微量滴定板杂交在取自42/48例无症状和53/56例有症状受试者的样本中检测到HIV-1序列。在90个样本中观察到两种检测方法的结果一致，其中81次检测为阳性，9次检测为阴性。在对14个不一致样本进行重复评估时，9个样本显示一致的阳性结果，接近检测方法的检测限。对ACH-2细胞进行系列稀释后进行扩增，并使用微量滴定板检测法检测PCR产物，得到半定量结果。这种简单、快速检测方法的灵敏度与更繁琐的DNA检测系统相当。这可能成为HIV-1研究及血清阳性个体临床护理中的有用工具。
