MOLECULAR INVESTIGATION OF THE ECOLOGY OF EEE VIRUS

EEE 病毒生态学的分子研究

• 批准号: 3454206
• 负责人: Patricia M. Repik
• 金额: $ 9.44万
• 依托单位: ALLEGHENY UNIVERSITY OF HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-07-01 至 1992-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3454206
• 关键词: Aves; Chelonia; Culicidae; Mexico; affinity chromatography; biological polymorphism; centrifugation; chemical fingerprinting; chickens; complementary DNA; ecology; endonuclease; enzyme linked immunosorbent assay; epizootiology; gene expression; gene mutation; genetic strain; genome; immunoprecipitation; latent virus infection; microorganism population study; molecular cloning; molecular genetics; nucleic acid hybridization; nucleic acid probes; tissue /cell culture; virus RNA; virus protein; virus replication

This investigation proposes to delineate two critical elements in the ecology of Eastern equine encephalitis (EEE) virus: (1) the molecular variation exhibited by EEE viruses over time within several different geographic foci of infection, and (2) elucidation of the overwintering maintenance mechanism(s) through the novel approach of using molecular probes. EEE virus is a mosquito-borne alphavirus which is widely distributed throughout both North and South America. Disease occurs annually. Although major outbreaks are intermittent, infection results in serious disease with high fatality and significant sequelae in survivors. Much work has been done to describe the epidemiology of EEE virus, but many questions remain unanswered concerning the ecology of EEE viruses, e.g., the number of different EEE strains circulating in a focus at any one time, the genetic and antigenic variation between endemic and epidemic strains, and the vector(s) responsible for transmitting disease to equines and humans. Similarly, although many hypotheses exist for the overwintering maintenance mechanism(s) of EEE virus, all previous attempts to isolate infectious virus in order to delineate the insect or mammalian winter reservoir(s) have been unsuccessful. In the studies outlined, a combination of molecular methods will be used to identify and characterize EEE viruses isolated from several different geographic foci over time. Characterization of viral RNA by oligonucleotide fingerprinting, hybridization, and restriction fragment analysis of virus-specific cDNA, and analysis of viral proteins by heterologous immunoprecipitation and enzyme digest analyses will enable us to define the degree of genetic and/or phenotypic differences occurring among viruses circulating in several natural endemic foci. Using this information, molecular probes will be developed for specific and rapid identification purposes. Finally, these probes will be used to directly examine tissues of suspected vertebrate hosts and invertebrate vectors for the presence of latent or persistent virus infections, in order to define to elusive overwintering maintenance mechanism(s) of EEE viruses. Once these aspects of EEE virus ecology have been determined, it may be possible to implement more effective surveillance and control strategies to curb outbreaks of this debilitating and highly fatal disease. This information may also serve as a model for defining the ecology of other important arboviruses.

这项调查建议描述两个关键要素 东马脑炎（EEE）病毒的生态学：（1） EEE病毒随着时间的流逝而表现出的分子变异 感染的几个不同地理焦点，（2）阐明 通过小说的越冬维护机制 使用分子探针的方法。 EEE病毒是一种蚊子传播的α病毒，广泛 分布在北美和南美。 疾病 每年发生。 尽管重大暴发是间歇性的，但 感染导致严重的疾病，死亡高， 幸存者中的明显后遗症。 已经做了很多工作 描述EEE病毒的流行病学，但许多问题 关于EEE病毒的生态，例如 在任何焦点上流通的不同EEE菌株的数量 一次，流行之间的遗传和抗原变异 和流行病，以及载体负责 向马和人类传播疾病。 同样，虽然 越冬维护存在许多假设 EEE病毒的机制，以前的所有试图分离 传染病为了描绘昆虫或哺乳动物 冬季水库没有成功。 在概述的研究中，分子方法的组合将 被用来识别和表征从与之隔离的EEE病毒 随着时间的流逝，几个不同的地理焦点。 表征 通过寡核苷酸指纹，杂交和 病毒特异性cDNA的限制片段分析和分析 通过异源免疫沉淀和酶的病毒蛋白质 消化分析将使我们能够定义遗传程度 和/或表型差异在循环的病毒中发生 在几个自然流行焦点中。 使用此信息， 分子探针将针对特定和快速开发 识别目的。 最后，这些探针将用于 直接检查可疑脊椎动物宿主的组织和 存在潜在或持续病毒的无脊椎动物载体 感染，以定义难以捉摸的越冬 EEE病毒的维护机制。 一旦这些方面 已经确定了EEE病毒生态学，可能有可能 实施更有效的监视和控制策略 遏制这种衰弱和高度致命疾病的暴发。 这 信息还可以作为定义生态的模型 其他重要的arbovirus。