Morandi P A, Schockmel G A, Yerly S, Burgisser P, Erb P, Matter L, Sitavanc R, Perrin L
Laboratory of Virology and AIDS Center, Geneva University Hospital, Switzerland.
J Clin Microbiol. 1998 Jun;36(6):1534-8. doi: 10.1128/JCM.36.6.1534-1538.1998.
A total of 234 pools were prepared from 10,692 consecutive serum samples negative for antibodies to human immunodeficiency virus type 1 (HIV-1) and HIV-2 collected at five virological laboratories (average pool size, 45 serum samples). Pools were screened for the presence of HIV-1 RNA by a modified commercial assay (Amplicor HIV-1 Monitor test) which included an additional polyethylene glycol (PEG) precipitation step prior to purification of viral RNA (PEG Amplicor assay). The sensitivity of this assay for HIV-1 RNA detection in individual serum samples within pools matches that of standard commercial assays for individual serum samples, i.e., 500 HIV-1 RNA copies per ml. Five pools were identified as positive, and each one contained one antibody-negative, HIV-1 RNA-positive serum sample, corresponding to an average of 1 infected sample per 2,138 serum samples. Retrospective analysis revealed that the five HIV-1 RNA-positive specimens originated from individuals who had symptomatic primary HIV-1 infection at the time of sample collection and who were also positive for p24 antigenemia. We next assessed the possibility of performing the prepurification step by high-speed centrifugation (50,000 x g for 80 min) of 1.5-ml pools containing 25 microl of 60 individual serum samples, of which only 1 contained HIV-1 RNA (centrifugation Amplicor assay). The sensitivity of this assay also matches the sensitivities of standard commercial assays for HIV-1 RNA detection in individual serum samples. The results demonstrate that both assays with pooled sera can be applied to the screening of large numbers of serum samples in a time- and cost-efficient manner.
从五个病毒学实验室收集的10,692份连续的1型人类免疫缺陷病毒（HIV-1）和2型人类免疫缺陷病毒抗体阴性的血清样本中，共制备了234个混合样本（平均混合样本大小为45份血清样本）。通过一种改良的商业检测方法（Amplicor HIV-1 Monitor检测）对混合样本进行HIV-1 RNA筛查，该方法在纯化病毒RNA之前增加了一个聚乙二醇（PEG）沉淀步骤（PEG Amplicor检测）。该检测方法对混合样本中单个血清样本中HIV-1 RNA检测的灵敏度与标准商业检测方法对单个血清样本的灵敏度相当，即每毫升500个HIV-1 RNA拷贝。鉴定出5个混合样本为阳性，每个样本都包含1份抗体阴性、HIV-1 RNA阳性的血清样本，相当于每2,138份血清样本中平均有1个感染样本。回顾性分析显示，这5个HIV-1 RNA阳性标本来自样本采集时患有症状性原发性HIV-1感染且p24抗原血症也呈阳性的个体。接下来，我们评估了对含有25微升60份个体血清样本（其中只有1份含有HIV-1 RNA）的1.5毫升混合样本进行高速离心（50,000×g，80分钟）以进行预纯化步骤的可能性（离心Amplicor检测）。该检测方法的灵敏度也与标准商业检测方法对单个血清样本中HIV-1 RNA检测的灵敏度相当。结果表明，这两种混合血清检测方法都可以以省时且经济高效的方式应用于大量血清样本的筛查。
