ENHANCED SRV/D DIAGNOSIS AND CONTROL

增强的 SRV/D 诊断和控制

• 批准号: 2284129
• 负责人: NICHOLAS WILLIAM LERCHE
• 金额: $ 17.37万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-12-01 至 1996-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2284129
• 关键词: Macaca fascicularis; Macaca mulatta; Macaca nemestrina; Retroviridae disease; active immunization; animal colony; communicable disease control; communicable disease diagnosis; diagnosis design /evaluation; epizootiology; laboratory rabbit; latent virus infection; polymerase chain reaction; recombinant proteins; simian virus; synthetic peptide; synthetic vaccines; type D retrovirus group; viral vaccines; virus envelope; virus protein

The objective of this three year proposal is to continue development and application of enhanced serological and virological tests for detection of simian type D retrovirus (SRV/D) infection in macaques, and to evaluate the potential role of recombinant vaccines in a comprehensive strategy for SRV/D control and eradication. We propose to continue our development of the polymerase chain reaction (PCR) as a diagnostic tool for SRV/D detection using both "generic" and virus type-specific primers and probes. The potential of PCR to replace more expensive and time consuming viral isolation methods will be evaluated by direct comparison of antibody detection, virus isolation and PCR in clinical specimens. The generic and type specific capabilities o various sets of primers and probes will be validated against known SRV/D serotypes, as well as against uncharacterised field isolates. In addition, we will develop and utilize "second generation" immunoassays for SRV/D antibody detection using recombinant viral proteins and synthetic peptides, with the objective of bringing on-line diagnostic assays with increased sensitivity and specificity. Tests with increased sensitivity are needed to streamline testing algorithms, and to reduce the frequency of "indeterminate" results. The performance of these recombinant assays will be compared with current assays that utilize disrupted whole virus as target antigen, against an extensive panel of well characterized antisera (positive, negative and indeterminate). Once developed and validated, PCR and recombinant antibody assays will be applied to screening and surveillance for development and maintenance of SPF breeding colonies of macaques. The use of improved diagnostic methods in testing strategies could potentially accelerate the rate of SPF colony development significantly. There is evidence that SRV/D infection in macaques is "vaccine preventable", yet the use of vaccines as part of an over-all program of control and eradication has not, to date, been given serious consideration. We propose to extend previous studies to answer some existing questions regarding vaccine efficacy directly relevant to their potential role in infection control strategies. Two major question will be addressed, 1) are these recombinant vaccines protective under conditions of natural exposure? 2) do these vaccines protect against establishment of latent infections? To achieve these objectives we will carry out a vaccine trial by housing immunized and sham immunized juvenile rhesus macaques together with infecteds, as a group in a corn-crib, essentially recreating the type of situation where SRV/D is most highly infectious. The vaccine efficacy will be determined by clinical, virological and serological monitoring, including PCR. An efficacious vaccine could prove to be a valuable adjunct to serological and virological screening programs in the control of SRD/D in macaques.

这三年提案的目的是继续发展， 应用血清学和病毒学测试的应用以检测 猕猴中的D型D型逆转录病毒（SRV/D）感染，并评估 重组疫苗在综合策略中的潜在作用 SRV/D控制和消除。我们建议继续我们的发展 聚合酶链反应（PCR）作为SRV/D的诊断工具 使用“通用”和病毒类型特异性引物和探针检测。 PCR取代更昂贵和耗时的病毒的潜力 隔离方法将通过直接比较抗体进行评估 临床标本中的检测，病毒分离和PCR。通用和 类型特定功能o各种引物和探针将是 针对已知的SRV/D血清型验证，并针对 未表征的场分离株。此外，我们将开发和利用 使用SRV/D抗体检测的“第二代”免疫测定 重组病毒蛋白和合成肽，目的 以提高灵敏度和 特异性。需要提高灵敏度的测试才能简化 测试算法，并降低“不确定”的频率 结果。这些重组测定的性能将与 利用整个病毒作为靶抗原的当前测定法 与广泛特征良好的Antisera面板（正面， 负和不确定）。一旦开发和验证，PCR和 重组抗体测定将应用于筛查和监视 用于开发和维护猕猴的SPF育种菌落。这 在测试策略中使用改进的诊断方法可能有可能 加速了SPF菌落发展的速度。 有证据表明，猕猴中的SRV/D感染是“疫苗 可预防的”，但使用疫苗作为全部计划的一部分 迄今为止，尚未对控制和根除 考虑。我们建议扩展以前的研究以回答一些 有关疫苗功效的现有问题与他们的疗效直接相关 在感染控制策略中的潜在作用。两个主要问题将是 解决，1）在条件下这些重组疫苗保护性是 自然暴露？ 2）这些疫苗可以防止建立 潜在感染？为了实现这些目标，我们将进行疫苗 通过住房免疫和假免疫的少年恒河猕猴试验 与受感染者一起，作为玉米订阅中的一组，本质上是重新创建的 SRV/D最感染力的情况类型。疫苗 功效将由临床，病毒学和血清学决定 监测，包括PCR。有效的疫苗可能被证明是 在血清学和病毒学筛查计划中有价值的辅助 猕猴中的SRD/D控制。