MOLECULAR DIAGNOSTIC TESTS FOR SIMIAN HERPESVIRUSES

描述（由申请人提供）：检测和诊断的能力 非人类灵长类病毒的感染远未达到最佳效果， 根本不存在 随着无特定病原体猴群的发展， 必须对猴病毒进行最高灵敏度的检测， 可以开发和监控这些殖民地。 的总体目标 拟议的研究是开发猕猴BV的基本分子数据， 为了利用这些结果来开发更灵敏的诊断测试， 比现有的更安全、更有效。 实现这些 目标，将进行以下实验：1）指示细胞系 将构建用于在表达GFP的病毒分离中的诊断用途 当感染BV时，使BV的病毒分离检测更加敏感， 更安全的程序。2)将获得BV（ABV）的致死缺失突变株， 通过删除必需的结构基因并构建 补充细胞系，其中ABV可以复制。 该系统（ABV + 补充细胞系）将使BV抗原和DNA的制备更加方便， 更安全3)将通过评估猕猴来鉴定主要BV免疫原 免疫应答的个体BV蛋白。 这些信息将使 开发有效的疫苗和重组抗原， 诊断测试4)BV的特异性糖蛋白基因将使用 诱导型哺乳动物表达系统，以及用于 发展免疫分析。 5)将开发一种TaqMan检测方法， 并区分单纯疱疹病毒-1（HSV I）、单纯疱疹病毒-2 （HSV 2）和BV。 这将提供一种极其灵敏的诊断方法 人类BV病例。 成功完成拟议的实验将 提供了一些试剂，可以更安全地检测 临床样本中感染性BV，BV抗原制备更安全， 诊断测试，更敏感和特异的血清学检测 猕猴和HSV阳性人血清中的抗BV抗体， 关于BV抗原决定簇的信息， 对感染的保护性免疫。

DESCRIPTION (Provided by applicant): The capability to detect and diagnose infections by non-human primate viruses is far less than optimal and in cases non-existent. As Specific Pathogen Free monkey colonies are developed, it will be essential that maximally sensitive tests for monkey viruses are available to develop and monitor such colonies. The overall goal of the proposed research is to develop basic molecular data for BV of macaques and to utilize these results to develop diagnostic tests that are more sensitive, more specific and safer than those currently available. To achieve these goals, the following experiments will be performed: 1) An indicator cell line will be constructed for diagnostic use in virus isolation which expresses GFP when infected by BV, making virus isolation testing for BV a more sensitive and safer procedure. 2) A lethal-deletion mutant strain of BV (ABV) will be constructed by deleting an essential structural gene and constructing a complementing cell line in which ABV can replicate. This system (ABV + complementing cell line) will make preparation of BV antigens and DNA much safer. 3) Major BV immunogens will be identified by assessing the macaque immune response to individual BV proteins. This information will allow development of effective vaccines and recombinant antigens for use in diagnostic tests. 4) Specific glycoprotein genes of BV will be expressed using an inducible mammalian expression system, and the expressed proteins used to develop immunciassays. 5) A TaqMan assay will be developed that can detect and differentiate herpes simplex virus-1 (HSV I), herpes simplex virus-2 (HSV2) and BV. This will provide an extremely sensitive means of diagnosing human BV cases. Successful completion of the proposed experiments will provide a number of reagents that will allow much safer detection of infectious BV in clinical samples, safer preparation of BV antigen for diagnostic tests, more sensitive and specific serological tests for detection of anti-BV antibodies in both macaque and HSV-positive human sera, and basic information regarding the antigenic determinants of BV which may induce protective immunity against infection.