MOLECULAR DIAGNOSTIC TESTS FOR SIMIAN HERPESVIRUS

猴子，特别是猕猴，是人类生存的重要资源 生物医学研究。猴疱疹病毒(B病毒)是一种自然病原体 猕猴，对工作的生物医学人员来说是一个非常真实的危险 和这些动物在一起。检测受感染猴子的敏感试验 需要识别对人员构成危险的动物，并 鉴定无B病毒动物用于建立无B病毒动物 繁育的殖民地。H.CerCopithicus 1(SA8)在狒狒中引起问题 繁育的殖民地。将SA8病毒传播给人类的发生率为 未知。能区分B病毒、SA8病毒的敏感试验 而两种人类单纯疱疹病毒(HSV)是 准确、快速地识别人类疱疹病毒感染。 灵长类疱疹病毒具有广泛的抗原性相关性 这是实现这一目标的主要障碍。以生成的数据为基础 从我们实验室的分子研究中，我们将开发出敏感的 灵长类病毒的酶联免疫吸附试验快速检测和鉴别 B病毒和SA8抗体。这些测试将基于特定的 病毒糖蛋白。 将建立灵长类疱疹病毒的聚合酶链式反应检测方法 临床样本。使用位于该基因保守区的一组引物 糖蛋白gB基因的一个病毒特异性区域的基因将 从四种灵长类疱疹病毒中的任何一种扩增出来的。后续 对聚合酶链式反应产物的分析可以确定哪种病毒 (HSV1、HSV2、B病毒或SA8)存在于临床样本中。ELISAs 还将开发用于疑似人类的血清学检测和 猴B病毒和/或SA8病例。ALL的GG糖蛋白同源物 有四种病毒具有抗原性。SA8和SA8的gG基因 BV将被克隆、测序并在哺乳动物细胞中稳定表达。 这些细胞系和抗GG抗体将用于开发 用于检测BV和SA8抗体的敏感的、病毒特异性的ELISA。 该项目将基础分子研究的成果应用于 B病毒和SA8病毒的检测和诊断是临床上重要的问题。 开发的分析方法将允许快速、灵敏和可靠 灵长类疱疹病毒感染的鉴别诊断。这将是 使灵长类群体管理者能够更有效地运营灵长类 设施和减少人畜共患疱疹造成的生命损失 感染。

Monkeys, particularly rhesus macaques, are a vital resource for biomedical research. Herpesvirus simiae (B virus), a natural pathogen of macaques, presents a very real danger for biomedical personnel who work with these animals. Sensitive assays for detection of infected monkeys are needed to identify animals which pose a risk to personnel and to identify B virus-free animals for use in establishment of B virus-free breeding colonies. H. cercopithicus 1 (SA8) causes problems in baboon breeding colonies. The incidence of transmission of SA8 to humans is unknown. Sensitive assays which can differentiate between B virus, SA8 and the two human herpes simplex viruses (HSVs) are necessary for accurate and rapid identification of human simian herpesvirus infections. The extensive antigenic relatedness of the primate herpesviruses has been a major impediment to achieving this goal. Building on data generated from molecular research in our laboratory, we will develop sensitive ELISA tests for rapid detection and differentiation of primate viruses and antibody to B virus and SA8. These tests will be based on specific viral glycoproteins. A PCR assay will be developed for detection of primate herpesviruses in clinical samples. Using a primer set located in conserved regions of the glycoprotein gB gene, a virus-specific region of the gene will be amplified from any of the four primate herpesviruses. Subsequent analysis of the PCR product will enable determination of which virus (HSV1, HSV2, B virus or SA8) is present in the clinical samples. ELISAs will also be developed for serological detection of suspected human and simian B virus and\or SA8 cases. The gG glycoprotein homologs of all four viruses are antigenically virus-specific. The gG genes of SA8 and BV will be cloned, sequenced and stably expressed in mammalian cells. These cell lines and anti-gG antibodies will be used to develop sensitive, virus-specific ELISAs for detection of antibody to BV and SA8. This project applies results of basic molecular research to the clinically important problem of B virus and SA8 detection and diagnosis. The assays developed will permit rapid, sensitive and reliable differential diagnosis of primate herpesvirus infections. This will enable primate colony managers to more effectively operate primate facilities and reduce loss of human life due to zoonotic herpes infections.