Simple monitoring of antiretroviral therapy with a signal-amplification-boosted HIV-1 p24 antigen assay with heat-denatured plasma
Simple monitoring of antiretroviral therapy with a signal-amplification-boosted HIV-1 p24 antigen assay with heat-denatured plasma
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DOI：
10.1097/00002030-199706000-00001
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发表时间：
1997-05-01
期刊：
影响因子：
3.8
通讯作者：
Schupbach, J
中科院分区：
文献类型：
--
作者：
Boni, J;Opravil, M;Schupbach, J
目的：病毒载量测定已成为艾滋病毒患者管理中不可或缺的一部分，但依赖于昂贵的低通量检测。我们评估了一种高度改进的HIV-1 p24抗原检测程序，包括热介导的免疫复合物解离和信号扩增增强酶联免疫吸附测定（ELISA），是否适用于抗逆转录病毒治疗监测。设计和方法：对23例CD4+ T细胞< 50 × 10(6)/l的患者在0、2、6、12、18、24、30和36周采集的127份血浆样本进行血浆病毒载量测定，这些患者在既往抗逆转录病毒治疗的基础上每天接受3次茚地那韦800 mg。检测包括病毒RNA的聚合酶链反应（PCR），使用罗氏扩增试剂盒进行前瞻性检测，以及使用杜邦HIV-1 p24 Core Profile ELISA与酪胺信号扩增步骤相结合，对热变性样品进行p24抗原的回顾性批量检测。颗粒相关逆转录酶（RT）通过产品增强RT （PERT）试验被确定为独立的第三方意见病毒载量标记物。结果：p24抗原检测与病毒RNA检测一样灵敏。在25周（0-39周）的中位观察时间内，抗原和RNA的总检出率分别为75.6%和73.6%。抗原检出限为0.2 pg/ml。在所有23个基线样本中均可检测到抗原，而在一个样本中未检测到RNA。79个样品中抗原和RNA水平均为阳性，相关系数r = 0.714 （P < 0.0001）。8个时间点p24抗原和RNA水平的平均变化相关系数r = 0.982 （P < 0.0001）。在个别患者中，这两个参数表现相似，在某些情况下几乎完全相同。所有样本的RT活性均可测量。结论：该抗原检测程序的性能可与RNA PCR相媲美，从而为监测抗逆转录病毒治疗的疗效提供了一种简单、高通量的替代方法。
Objective: Virus load determination has become indispensable for the management of HIV patients, but depends on expensive assays of a low throughput. We evaluated whether a highly improved HIV-1 p24 antigen detection procedure which involves heat-mediated immune complex dissociation and signal-amplification-boosted enzyme-linked immunosorbent assay (ELISA) was suitable for antiretroviral treatment monitoring.Design and methods: Virus load in plasma was determined for 127 plasma samples taken at 0, 2, 6, 12, 18, 24, 30 and 36 weeks from 23 patients with CD4+ T cells < 50 x 10(6)/l who received indinavir 800 mg three times daily in addition to prior antiretroviral treatment. Tests included polymerase chain reaction (PCR) for viral RNA, measured prospectively with the Roche Amplicor kit, and retrospective batch testing of heat-denatured samples for p24 antigen by the DuPont HIV-1 p24 Core Profile ELISA linked with a tyramide signal amplification step. Particle-associated reverse transcriptase (RT) by the product-enhanced RT (PERT) assay was determined as an independent third-opinion viral load marker.Results: p24 antigen was detected as sensitively as viral RNA. Overall detection during a median observation time of 25 weeks (range, 0-39) amounted to 75.6% for antigen and 73.6% for RNA. The antigen detection limit was 0.2 pg/ml. Antigen was detectable in all 23 baseline samples, whereas RNA was undetectable in one. Antigen and RNA levels in 79 samples positive for both markers correlated with r = 0.714 (P < 0.0001). Average changes in levels of p24 antigen and RNA at eight timepoints correlated with r = 0.982 (P < 0.0001). In individual patients, the two parameters behaved similarly, and in certain cases virtually identically. RT activity was measurable in all samples.Conclusions: The performance of this antigen detection procedure is comparable to RNA PCR, thus providing a simple, high throughput alternative in monitoring the efficacy of antiretroviral treatment.