RAPID HOMOGEONEOUS HEMAGGLUTINATION ASSAY FOR HIV

HIV 快速均质血凝测定

• 批准号: 6170866
• 负责人: JAMES W LARRICK
• 金额: $ 35.75万
• 依托单位: PANORAMA RESEARCH, INC.
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-03-01 至 2001-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6170866
• 关键词: HIV envelope protein gp41; bacteriophage M13; biomarker; communicable disease diagnosis; diagnosis design /evaluation; glycophorin; hemagglutination test; human immunodeficiency virus 1; human tissue; rapid diagnosis; recombinant virus; technology /technique development; virus protein

The basic idea is to create a homogenous phage hemagglutination assay (HPA-homogeneous phage agglutination) for the rapid diagnosis of HIV infection. The design employs M13 phages displaying two different molecules on their surface, an antibody or other binding site able to react with a conserved cell surface marker of human erythrocytes (e.g., glycophorin A), fused to protein 3, and an antigen representative of a human infectious agent, fused to protein 8. In phase I, we created and validated this assay using an HIV-1 gp41 peptide with proven diagnostic value. Recombinant phage particles placed in direct contact with red cells and a positive control antisera induced erythrocyte agglutination indicating that the anti-HIV antigen antibodies present in the serum bridged the phage particles attached to different red blood cells. Importantly, negative control antisera did not show agglutination. Based on these positive results, the goals of phase II are to 1) Optimize the assay using human sera panels and/or blood from infected individuals; 2) Scale-up the diagnostic phage particles, and prepare a clinical prototype diagnostic reagent kit ; 3) establish assay variability, quality control, etc. Finally, we will evaluate performance of this Rapid HIV Homogeneous Phage Assay on clinical samples in collaboration with the Division of AIDS, STDs and Tuberculosis Laboratory Research at the CDC. PROPOSED COMMERCIAL APPLICATIONS: Development of a rapid HIV homogeneous assay will have significant commercial potential for the field and office diagnosis of HIV.

基本思想是创建一种均质噬菌体血凝测定（HPA-均质噬菌体凝集），用于快速诊断 HIV 感染。该设计采用在其表面展示两种不同分子的 M13 噬菌体、一种能够与人类红细胞保守细胞表面标记物（例如血型糖蛋白 A）反应的抗体或其他结合位点（与蛋白 3 融合）以及代表人类传染原的抗原（与蛋白 8 融合）。在第一阶段，我们使用 HIV-1 gp41 肽创建并验证了该测定法 已证实的诊断价值。重组噬菌体颗粒与红细胞和阳性对照抗血清直接接触，诱导红细胞凝集，表明血清中存在的抗HIV抗原抗体桥接了附着于不同红细胞的噬菌体颗粒。重要的是，阴性对照抗血清没有表现出凝集。基于这些积极结果，第二阶段的目标是 1) 使用人类血清组和/或感染个体的血液优化检测； 2）放大诊断噬菌体颗粒，制备临床原型诊断试剂盒； 3) 建立测定变异性、质量控制等。最后，我们将与 CDC 艾滋病、性传播疾病和结核病实验室研究部门合作，评估这种快速 HIV 均质噬菌体测定在临床样本上的性能。拟议的商业应用：快速 HIV 均相检测的开发对于 HIV 的现场和办公室诊断具有巨大的商业潜力。