DETECTION, QUANTITATION & CHARACTERIZATION OF HIV

检测、定量

• 批准号: 6436551
• 负责人: INDIRA HEWLETT
• 金额: --
• 依托单位: BUREAU OF HEALTH PLANNING AND RESOURCES DEVELOPMENT
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6436551
• 关键词: AIDS /HIV test; HIV infections; Parvoviridae; blood chemistry; clinical research; diagnosis design /evaluation; disease /disorder proneness /risk; epidemiology; hepatitis B virus group; hepatitis C virus; human immunodeficiency virus 1; human immunodeficiency virus 2; human tissue; nucleic acid probes; nucleic acid sequence; polymerase chain reaction; virus RNA; virus genetics; virus load

1) A plasma specimen with a high titer of HIV RNA was identified and tested for use as the first generation CBER standard. A second standard based on culture supernatant from infected cells (clade B) was developed. Both materials have been evaluated in international collaborative studies involving the NIBSC and WHO for assessment of assay performance. The plasma specimen in a lyophilized form has been accepted as the first in international WHO standard. Currently, the laboratory is focussing on developing an HIV-1 subtype panel. Two isolates from each clade have been cultured to high titers. In a limited collaborative study, the copy numbers were estimated in the samples by the TMA, PCR and bDNA techniques. These reagents will be useful in regulatory and global assay standardization efforts. 2) In-house serologic and genetic assays and Taqman probes are being developed for HIV-1 group O and non clade B subtypes. An in-house HIV group O ELISA and primers and probes for HIV group O and M have been developed. The probes will be used to generate microarray based assays for viral genotype. These assays will serve as tools in molecular epidemiologic studies to determine prevalence of non clade B strains particularly HIV-1 group O in endemic areas. We have recently initiated a study to study molecular evolution of HIV in Camerooon a region which harbors several subtypes. We are also comparing the performance of various direct viral marker assays such as PERT, RT-PCR, and p24 assays for their ability to detect viral variants and HIV in the early phase of infection.

1）鉴定出具有高滴度HIV RNA的血浆标本并测试以用作第一代CBER标准。 开发了基于感染细胞（进化枝B）的培养上清液的第二个标准。 两种材料均已在涉及NIBSC和评估测定性能评估的国际合作研究中进行了评估。冻干形式的等离子体标本已被接受为国际标准的第一个标准。目前，该实验室的重点是开发HIV-1亚型面板。每个进化枝中的两个分离物已被培养为高滴度。在一项有限的协作研究中，TMA，PCR和BDNA技术在样品中估算了拷贝数。这些试剂将在监管和全球测定标准化工作中有用。 2）为HIV-1组O和非进化枝B亚型开发了内部血清学和遗传测定和Taqman探针。 已经开发了一个内部的HIV hiv组O ELISA以及HIV组O和M的引物和探针。这些探针将用于生成基于微阵列的病毒基因型测定法。这些测定将用作分子流行病学研究的工具，以确定非进化核B菌株的患病率，尤其是在流行地区的HIV-1组O。我们最近开始了一项研究，研究了摄影室中HIV的分子进化，该区域具有多种亚型。我们还比较了各种直接病毒标记物测定的性能，例如PERT，RT-PCR和P24分析，以便在感染的早期检测病毒变异和HIV的能力。