VIROLOGY DEVELOPMENT RESEARCH

HIV-1感染有效治疗方法的发展与评价 受限于到临床终点的较长时间间隔，以及 疾病的病毒学标志物相关性相对较差或缺乏 进步。婴幼儿慢性阻塞性肺疾病的科学调查与治疗 潜在的有毒抗病毒药物因以下事实而变得复杂 缺乏可靠的方法在第一时间诊断HIV-1感染 因被动转移母体HIV-1而存活数月 抗体。需要能够量化HIV-1病毒载量的病毒学检测 在患者的血液中进行实验性的抗病毒药物治疗 提供更早的药物疗效评估和2)准确检测 婴儿感染HIV-1。这项拟议研究的目的如下： 1.标准化验证敏感度、特异度和 商业聚合酶链式反应(PCR)检测方法的重复性 HIV-1序列的非同位素检测。 2.HIV-1前病毒DNA与游离HIV-1数量变化的相关性 用聚合酶链式反应和其他病毒学方法检测血浆中的RNA水平， 接受治疗的患者的CD4计数、临床疾病分期和临床结果 ACTG协议。 3.制定和评估标本处理程序和HIV-1聚合酶链式反应检测 用于外周血中HIV-1前病毒DNA序列的非同位素检测 外周血单个核细胞(PBMC)、脐血单个核细胞(CBMC)和 HIV-1血清阳性母亲新生儿的干血斑点。 为了实现这些目标，HIV-1的血液样本呈阳性 成人和新生儿将从一个或多个ACTU站点收集，以及 为HIV-1聚合酶链式反应检测和定量终点进行前瞻性处理 稀释法。HIV-1聚合酶链式反应批量检测将使用 Perkin-Elmer/GenProbe HIV-1聚合酶链式反应检测和/或罗氏HIV-1 聚合酶链式反应检测方法。在初步评估HIV-1聚合酶链式反应的敏感性后 和特异性，定性和定量HIV-1聚合酶链式反应检测 与临床结果、CD_4计数和其他病毒学检测相关 (HIV-1培养，血清p24抗原水平)，将进行前瞻性评估 在特定的儿科和成人ACTG方案内。

The development and evaluation of effective therapies for HIV-1 infection is limited by the long time interval to clinical endpoints and the relatively poor correlation or absence of virological markers for disease progression. Scientific investigation and treatment of infants with potentially toxic antiviral drugs are complicated by the fact that there is a lack of reliable methods to diagnose HIV-1 infection within the first several months of life due to passive transfer of maternal HIV-1 antibodies. Virological assays are needed that can 1) quantify HIV-1 load in the blood of patients on experimental antiviral drug therapies thereby providing an earlier assessment of drug efficacy and 2) accurately detect HIV-1 infection in infants. The proposed study has the following aims: 1. Standardize and validate the sensitivity, specificity, and reproducibility of commercial polymerase chain reaction (PCR) assays for the nonisotopic detection of HIV-1 sequences. 2. Correlate quantitative changes in HIV-1 proviral DNA and cell free HIV-1 RNA levels in plasma, as measured by PCR, with other virological assays, CD4 counts, clinical stage of disease, and clinical outcomes in patients on ACTG protocols. 3. Develop and evaluate specimen processing procedures and HIV-1 PCR assays for the nonisotopic detection of HIV-1 proviral DNA sequences in peripheral blood mononuclear cells (PBMC), cord blood mononuclear cells (CBMC) and dried blood spots of newborns of HIV-1 seropositive mothers. In order to achieve these aims, blood specimens from HIV-1 seropositive adults and newborns will be collected from one or several ACTU sites, and processed prospectively for HIV-1 PCR detection and quantitative endpoint dilution assays. HIV-1 PCR batch testing will be performed using the Perkin-Elmer/Gen-Probe HIV-1 PCR detection assays and/or the Roche HIV-1 PCR detection assay. After the initial evaluation of HIV-1 PCR sensitivity and specificity, the ability of qualitative and quantitative HIV-1 PCR to correlate with clinical outcome, CD4 counts, and other virological assays (HIV-1 culture, serum p24 antigen levels), will be evaluated prospectively within specific pediatric and adult ACTG protocols.