Viral etiology of idiopathic chronic diarrhea in rhesus macaques

 描述（由申请方提供）：直到最近，尝试鉴定病毒病原体的研究仅限于检测通过细胞培养、PCR、抗原特异性抗体染色或通过血清转化可检测的先前已知病毒。因此，检测仅限于已经表征的病毒，主要是那些通过对体外生长细胞的致细胞病变作用发现的病毒。高通量DNA测序的引入现在允许宏基因组方法鉴定存在的所有病毒，并揭示了大量以前未知的病毒，属于许多不同的病毒家族，特别是在人类粪便以及野生和驯养动物中。核酸检测的高度敏感性也揭示了肠道病毒感染的高比率，不仅在健康的人类和动物中，而且在健康的人类和动物中。在这里，将使用宏基因组学/生物信息学方法首先生成存在于患有特发性慢性腹泻（ICD）的恒河猴的大队列中的所有肠道病毒的综合列表，所述特发性慢性腹泻（ICD）是类似于人类溃疡性结肠炎的结肠的常见炎性病症。为了确定与这种不明原因疾病相关的候选病毒病原体，经典病例对照研究将比较大量ICD动物与健康对照动物中存在的所有病毒的存在和病毒浓度。然后将使用原位杂交检测这些病毒在受ICD影响的动物组织病理学解剖部位的病毒复制。然后将最大数量的动物中与ICD最强相关的最佳候选病毒病原体经口接种至幼龄猕猴中，并密切监测结肠中ICD的发展、病毒脱落和病毒复制。因此，我们将应用宏基因组学和生物信息学工具的力量，结合经典的流行病学研究，对影响恒河猴研究群体的常见和慢性灵长类疾病中病毒的作用进行全面测试。兽医科学专家将仔细选择大量动物样本，并对其病毒核酸进行全面分析，然后对动物进行接种，以确定一种或多种病毒病原体。这项研究还将推进我们对大型灵长类动物群体中病毒多样性的理解，并证明宏基因组学在识别所有病毒方面的实用性，以用于随后的疾病关联研究，比较受影响病例与健康对照的病毒复制。导致ICD的病毒的鉴定不仅有助于管理生物医学研究中最常用的非人灵长类动物物种的健康，而且还将为病毒诱导的慢性腹泻/结肠炎提供模型系统，并为疫苗开发提供病毒靶点，以控制灵长类动物中心死亡率和动物变异性的主要来源。

 DESCRIPTION (provided by applicant): Until recently, studies attempting to identify viral pathogens were restricted to testing for previously known viruses detectable by cell culture, PCR, staining with antigen-specific antibodies, or through sero-conversion. Testing was therefore restricted to already characterized viruses, largely those that had been discovered through their cytopathic effects on cells grown in vitro. The introduction of high-throughput DNA sequencing now allows metagenomic approaches to identify all the viruses present and has revealed a large number of previously unknown viruses, belonging to many different viral families, particularly in the feces of humans as well as in wild and domesticated animals. The very high sensitivity of nucleic acid testing has also revealed a high rate of enteric viral infections not just in diarrheic but also in healthy humans and animals. Here a metagenomics/bioinformatics approach will be used to first generate a comprehensive list of all the enteric viruses present in a large cohort of rhesus macaques with idiopathic chronic diarrhea (ICD), a common inflammatory condition of the colon resembling human ulcerative colitis. In order to identify candidate viral pathogens associated with this unexplained disease, a classic case control study will compare the presence and viral concentration of all the virus present in a large number of animals with ICD versus healthy controls. Viral replication of these viruses in the anatomical site of tissue pathology in animals affected with ICD will then be tested using in situ hybridization. The top candidate viral pathogens, most strongly associated with ICD in the largest number of animals, will then be orally inoculated into juvenile macaques and the development of ICD, viral shedding, and viral replication in the colon closely monitored. We will therefore apply the power of metagenomics and bioinformatics tools combined with a classic epidemiological study for an all-inclusive test of the role of viruses in a common and chronic primate disease affecting rhesus macaque research colonies. A large numbers of animal samples will be carefully selected by veterinary science experts and their viral nucleic acids comprehensively analyzed followed by animal inoculations to identify one or more viral pathogens. This study will also advance our understanding of viral diversity in a large primate population, and demonstrate the utility of metagenomics to identify all viruses present for subsequent disease association studies comparing viral replication in affected cases versus healthy controls. The identification of virus(es) causing ICD will not only assist managing the health of the non-human primate species most often used in biomedical research but will also provide model systems for virally induced chronic diarrhea/colitis and viral targets for vaccine development to control a major source of mortality and animal variability in primate centers.