USE OF PCR FOR DIAGNOSING HIV-1 INFECTION IN CHILDREN

在该补助金期间进行的研究旨在建立 PCR检测在儿童HIV-1诊断中的意义 感染和疾病最终进展的预测。 一个简单的， 快速，可靠的检测适合于常规临床试验将是 开发并评估用于直接检测HIV-1序列， 血清阳性婴儿 本试验将包括聚合酶链反应 (PCR)扩增HIV-1基因组的两个区域， 寡聚体捕获测定以微量滴定孔形式进行。 的特异性和 将在实验室质控品上评价该检测试剂盒的灵敏度 标本，并选择最佳格式。 完成PCR的程序 临床样品的测试，包括样品制备，扩增， 和检测将被简化，以减少处理和培养时间。 将提供适当的对照品以验证试验结果。 将设计对检测试剂盒的其他修改，以允许检测 的HIV-1 RNA序列，并定量PCR结果。 RNA测试可以 作为活动性感染的有用指标， 疾病的进展。 定量测定将允许测量 病毒载量，并可用于监测对抗逆转录病毒的反应 治疗 在这项资助的过程中开发的测试将允许 对HIV-1感染婴儿进行更有意义的诊断， 感染管理。

Studies conducted in the course of this grant are aimed at establishing the significance of PCR based assays for the diagnosis of pediatric HIV-1 infection and prediction of eventual progression to disease. A simple, rapid, reliable assay suitable for routine clinical testing will be developed and evaluated for the direct detection of HIV-1 sequences in seropositive babies. This assay will consist of polymerase chain reaction (PCR) amplification of two regions of the HIV-1 genome coupled with an oligomer capture assay in a microtiter well format. The specificity and sensitivity of this assay will be evaluated on laboratory control specimens, and an optimum format selected. The procedures for complete PCR testing of clinical samples including sample preparation, amplification, and detection will be streamlined to reduce handling and incubation times. Appropriate controls will be provided to validate the results of the assay. Additional modifications to the assay will be designed to permit detection of HIV-1 RNA sequences and to quantitate the PCR results. The RNA test may serve as a useful indicator of active infection and may be predictive of progression of disease. The quantitative assay will permit measurement of viral load and be useful for monitoring the response to anti-retroviral treatment. The tests developed in the course of this grant will permit more meaningful diagnosis of HIV-1 infected babies and allow better management of infection.