Detection of human immunodeficiency virus type 1 (HIV-1) RNA in pools of sera negative for antibodies to HIV-1 and HIV-2
Detection of human immunodeficiency virus type 1 (HIV-1) RNA in pools of sera negative for antibodies to HIV-1 and HIV-2
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DOI：
10.1128/jcm.36.6.1534-1538.1998
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发表时间：
1998-06-01
影响因子：
9.4
通讯作者：
Perrin, L
中科院分区：
文献类型：
--
作者：
Morandi, PA;Schockmel, GA;Perrin, L
从5个病毒学实验室收集的10 692份人类免疫缺陷病毒1型（HIV-I）和HIV-2抗体阴性的连续血清样本中制备了共计234份样本池（平均样本池大小为45份血清样本）。通过改良的商业试验（Amplicor HIV-1 Monitor试验）筛选HIV-1 RNA的存在，该试验包括在纯化病毒RNA（PEG Amplicor试验）之前的额外聚乙二醇（PEG）沉淀步骤。该检测试剂盒检测合并液中单个血清样本中HIV-1 RNA的灵敏度与单个血清样本的标准商业检测试剂盒的灵敏度相匹配，即，每英里500个HIV-1 RNA拷贝。5个样本池被鉴定为阳性，每个样本池包含1份抗体阴性、HIV-1 RNA阳性血清样本，相当于平均每2，138份血清样本中有1份感染样本。回顾性分析显示，5份HIV-1 RNA阳性标本来自于样本采集时有症状性原发性HIV-1感染且p24抗原血症阳性的个体。接下来，我们评估了通过高速离心（50，000 x g，80分钟）1.5 ml合并液进行预纯化步骤的可能性，该合并液含有25 μ l的60份单独血清样本，其中仅1份含有HIV-1 RNA（离心Amplicor检测）。该检测试剂盒的灵敏度也与单个血清样本中HIV-1 RNA检测的标准商业检测试剂盒的灵敏度相匹配。结果表明，采用合并血清的两种检测试剂盒均可以以时间和成本有效的方式应用于大量血清样本的筛选。
A total of 234 pools were prepared from 10,692 consecutive serum samples negative for antibodies to human immunodeficiency virus type 1 (HIV-I) 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 mi. 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 mu l 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.