DEVELOPMENT AND EVALUATION OF NEW SCREENING TESTS

这项提案的目标和具体目的是开发改进的 检测HTLV-I、HTLV-II和HTLV-II抗体的方法 HIV-2抗原以及检测、表征和 对这些病毒的蛋白质和核酸进行量化。 所有的化验都将使用以前积累的和特征良好的 包含20,000多份人类血清和细胞库，以便于评估 敏感性和特异性。 血清学检测将被设计为检测免疫球蛋白或免疫球蛋白M 反应性。化验格式将涉及多种来源的病毒 抗原，包括纯化的病毒粒子、合成肽和重组 蛋白质。其目标将是开发酶联免疫吸附试验、免疫印迹和/或 足够敏感和特异的免疫印迹，以消除需要 给RIPA的。 蛋白质检测将集中在多克隆和 用于直接细胞染色或抗原的单抗 捕获化验格式。将针对所有主要的 结构、功能和调节的病毒蛋白。 核酸研究将集中在聚合酶链式反应的继续发展上 以及用于检测、定量和测序的QB复制酶系统 从人体组织和体液中提取特定的病毒DNA和RNA。 预计上述技术将进入 人类免疫缺陷病毒的临床预防、诊断和治疗系统 逆转录病毒病。

The objectives and specific aims of this proposal are to develop improved assays for both the detection of antibodies against HTLV-I, HTLV-II and HIV-2 antigens and also the detection, characterization, and quantification of the proteins and nucleic acids of these same viruses. All assays will make use of previously accrued and well-characterized bank of over 20,000 human sera and cells to facilitate assessment of sensitivity and specificity. Serological assays will be designed to detect either IgG or IgM reactivities. Assay formats will involve a variety of sources of viral antigen including purified virions, synthetic peptides and recombinant proteins. The aim will be to develop ELISA, western blots and/or immunoblots that are sensitive and specific enough to obviate the need for RIPA's. Protein detection will center on the development of polyclonal and monoclonal antibodies to be used in direct cell staining or antigen capture assay formats. Antibodies will be made against all major structural, functional, and regulatory viral proteins. Nucleic acid studies will center on the continued development of the PCR and QB replicase system for the detection, quantification and sequencing of specific viral DNA and RNA from human tissues and body fluids. It is anticipated that the above techniques will make their way into clinical systems for the prevention, diagnosis and treatment of human retroviral disorders.