使用热变性血浆的信号放大增强型HIV-1 p24抗原检测法对抗逆转录病毒疗法进行简单监测。

Simple monitoring of antiretroviral therapy with a signal-amplification-boosted HIV-1 p24 antigen assay with heat-denatured plasma.

作者信息

Böni J, Opravil M, Tomasik Z, Rothen M, Bisset L, Grob P J, Lüthy R, Schüpbach J

机构信息

Swiss National Centre for Retroviruses, University of Zurich, Switzerland.

出版信息

AIDS. 1997 May;11(6):F47-52. doi: 10.1097/00002030-199706000-00001.

DOI:10.1097/00002030-199706000-00001

PMID:9143600

Abstract

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.

摘要

目的

病毒载量测定已成为管理HIV患者不可或缺的手段，但依赖于昂贵且通量低的检测方法。我们评估了一种经过高度改进的HIV-1 p24抗原检测程序是否适用于抗逆转录病毒治疗监测，该程序涉及热介导的免疫复合物解离和信号放大增强酶联免疫吸附测定（ELISA）。

设计与方法

对23例CD4 + T细胞<50×10(6)/l且除接受过抗逆转录病毒治疗外还每日三次服用800 mg茚地那韦的患者，在第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。检测包括用罗氏Amplicor试剂盒前瞻性检测病毒RNA 的聚合酶链反应（PCR），以及通过与酪胺信号放大步骤联用的杜邦HIV-1 p24核心谱ELISA对热变性样本进行回顾性批量检测p24抗原。通过产物增强逆转录酶（PERT）测定法测定颗粒相关逆转录酶（RT），将其作为独立的第三方病毒载量标志物。

结果

p24抗原检测的灵敏度与病毒RNA相当。在中位观察时间25周（范围0 - 39周）内，抗原总体检测率为75.6%，RNA为73.6%。抗原检测下限为0.2 pg/ml。在所有23份基线样本中均可检测到抗原，而有一份样本未检测到RNA。79份两种标志物均呈阳性的样本中，抗原和RNA水平的相关性r = 0.714（P < 0.0001）。八个时间点p24抗原和RNA水平的平均变化相关性r = 0.982（P < 0.0001）。在个体患者中，这两个参数表现相似，在某些情况下几乎完全相同。所有样本中均可检测到RT活性。