APPLICATION OF GENETIC TECHNIQUES FOR DETECTION OF HIV IN CELLS AND BODY FLUIDS

1)允许检测和区分HIV-1和 通过聚合酶链式反应检测HIV-2病毒是为临床样本而设计的。我们有 还优化了检测HIV-1、-2、 HTLV-I，-II。该检测方法的性能已在临床上进行了评估。 混合感染患者的样本。 2)建立了一种检测病毒RNA的灵敏的RNA-PCR方法 在体液中。并应用于病毒的检测和定量 在患者的体液和感染细胞中。使用了这种方法 评估受感染者的唾液中是否存在艾滋病毒 商业产品。 3)一种新型的基于包被HIV的磁珠的病毒捕获方法 已开发出用于检测体内病毒RNA的环状病毒抗体 体液。该检测方法在HIV阳性患者中表现出很高的敏感性。 个人。一种非同位素格式也得到了优化。未来 应用包括对传染性“窗口期”的研究，这是一种主要的 关注艾滋病和艾滋病患者的血液安全和病毒载量评估 患者参与的抗病毒治疗。其中一项研究涉及一项 原发感染和进展性与非进展性感染的评估 病毒将通过聚合酶链式反应进行定量和表征的感染 病毒分离株的核苷酸序列测定和表型培养分析。 多种形式的检测也将针对艾滋病毒和丙型肝炎病毒进行优化。 用于窗口期检测的RNA。由于多路传输格式可以 实施这些技术的经济高效的方法 血液筛查，这些研究将使我们熟悉 在这类检测的优化和标准化方面。 4)正在努力制定艾滋病毒的量化标准 血浆中的核糖核酸。QC-聚合酶链式反应和定量聚合酶链式反应正在使用病毒库存进行评估 含有已知数量的HIV p24抗原的HIV呈阴性 血浆。这些试剂将与世界卫生组织合作进行评估 国际基因扩增技术工作组 HIV阳性患者血浆(Ag+ve/Ab+ve或-ve)中HIV RNA的定量 个人。

1) Primer pairs that permit detection and discrimination of HIV-1 and HIV-2 by PCR have been designed for use with clinical samples. We have also optimized a multiplex PCR assay for the detection of HIV-1, -2, HTLV-I, -II. The performance of the assay has been evaluated on clinical samples from patients with co-infections. 2) A sensitive RNA-PCR assay was developed for the detection of viral RNA in body fluids. and applied to the detection and quantitation of virus in body fluids and infected cells from patients. This method was used to assess the presence of HIV in saliva of infected individuals and in commercial products. 3) A novel, viral capture assay based on magnetic beads coated with HIV env antibodies has been developed for detection of viral RNA in body fluids. The assay has demonstrated high sensitivity in HIV positive individuals. A non isotopic format has also been optimized. Future applications include studies on the infectious "window" period, a primary concern to blood safety and virus load assessment in HIV disease and antiviral therapy involving patients. One of these studies involves an evaluation of primary infection and progressive vs. non progressive infection where virus will be quantitated by PCR and characterized by nucleotide sequencing of virus isolates and phenotypic culture assays. A multiplex format of the assay will also be optimized for HIV and HCV RNA for detection in the window phase. Since multiplex formats may be the cost-effective approach to implementation of these techniques for blood screening, these studies will allow us to be familiar with issues in optimization and standardization of such assays. 4) Efforts are in progress to generate standards for quantitation of HIV RNA in plasma. QC-PCR and Q-PCR are being evaluated using virus stocks containing known amounts of HIV p24 antigen spiked into HIV negative plasma. These reagents will be assessed in collaboration with the International Working Group on Gene Amplification techniques and used to quantitate HIV RNA in plasma (Ag+ve/Ab+ve or -ve) from HIV positive individuals.