Studies Of Gastroenteritis Viruses By Electron Microscopy

通过电子显微镜研究胃肠炎病毒

• 批准号: 8336041
• 负责人: Albert Z Kapikian
• 金额: $ 30.6万
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8336041
• 关键词: Annual Reports; Appearance; Area; Astrovirus; Australia; Baculoviruses; Calicivirus; Capsid Proteins; Cell Culture System; Centrifugation; Characteristics; Child; Classification; Clinical Research; Collaborations; Coronavirus; Detection; Dialysis procedure; Diarrhea; Disease; Electron Microscope; Electron Microscopy; Enteral; Epidemiologic Studies; Epidemiology; Etiology; Family; Feces; Gastroenteritis; Genotype; Goals; Gold; Hepatitis A Virus; Hepatitis E virus; Hepatitis Viruses; Hospitalized Child; Hospitals; Human; Imagery; Immune; Infant; Infantile Diarrhea; Infectious hepatitides; Influenza A Virus, H1N1 Subtype; Investigation; Laboratory Study; Mus; National Institute of Diabetes and Digestive and Kidney Diseases; Natural History; Norovirus; Norwalk virus; Odobenus rosmarus; Pediatrics; Play; Preparation; Research; Role; Rotavirus; Specimen; System; Techniques; Torovirus; Trefoil Motif; Vesivirus; Viral Gastroenteritis; Virion; Virus; Virus-like particle; ascorbate; cesium chloride; enteric adenovirus infection; influenzavirus; member; programs; tissue culture; tool

For over three decades, the electron microscope has been an integral part of the research program of the Epidemiology Section regarding the etiologic role, natural history and physical characteristics of various important enteric viruses. Past highlights of these investigations have included inquiry into the etiology of diseases of unknown cause involving studies of fastidious agents of disease that defied cultivation in any cell culture system. The electron microscope played a major role (i) in the discovery of the 27nm Norwalk virus (now known as a member of the norovirus genus in the calicivirus family) and the establishment of its etiologic role in human diarrheal disease, and (ii) in collaboration with the Hepatitis Viruses Section (HVS), in the discovery of the 27nm hepatitis A virus particle and establishment of its etiologic role in infectious hepatitis and in addition in studies of the hepatitis E virus with the HVS . It also was used as an important epidemiologic tool for the detection of rotaviruses in stools of infants and young children hospitalized with diarrhea and helped in establishing the importance of these agents in severe diarrhea of infants and young children. Although intensive efforts have been made, the Norwalk and related human enteric noroviruses have yet to be cultivated in any tissue culture system. This, along with the visualization of virus-like particles (VLPs) of certain members of this group of agents, have contributed to the continuing important role of the electron microscope in the research program of the LID. The electron microscope has continued to be an important primary and adjunctive tool in various capacities. Since last year's annual report, studies have included a variety of areas: (1) visualization of full walrus vesivirus VLPs in a purified preparation following dialysis; (2) visualization of Australian calicivirus VLPs from 2001, 2007 and 2008 (collaboration with Ms. Mahers and Dr. Kirkwood, Pediatrics, Royal Childrens Hospital, Australia); (3) visualization of chimeric VLPs of Norwalk virus shell and JHH3 P (collaboration with JHU); (4) attempt to determine if ascorbate has any direct effect on rotavirus SA-11 morphologically (collaboration with Dr. Levine of NIDDK) ; (5) attempts to visualize hepatitis E virus by direct EM, by IEM without centrifugation and IEM with centrifugation (collaboration with Dr. Emerson); (6) attempt to determine if hepatitis E virus genotype is genotype specific by IEM; (7) visualization of Norwalk virus- shell murine norovirus P domain chimeric capsid proteins expressed in baculovirus system;( 8) attempt to visualize morphologic appearance of influenza virus H1N1 by direct EM (collaboration with Dr. Lackdawala and Ms. Lamirande); (9) visualization of various murine norovirus preparations in CsCl ;(10) visualization of presence and integrity of VLP preparation of norovirus JHH3 being prepared for collaborative cryo- EM study at RML.

在过去的三十年中，电子显微镜一直是有关各种重要肠道病毒的病因学作用，自然历史和物理特征的流行病学部分研究计划的组成部分。这些研究的过去亮点包括对未知原因疾病的病因进行调查，这些病因涉及对任何细胞培养系统中违反培养的疾病的研究。电子显微镜在发现27nm Norwalk病毒（现在被称为钙病毒家族中诺如病毒属的成员）中起着重要作用（i），并在人类腹泻病中建立了病因学作用，以及（ii）与肝炎病毒病毒的合作（II），在肝炎病毒部分的合作（HVS）中的发现（Hepaties）在感染性肝炎中的作用以及HVS的丙型肝炎病毒的研究。它也被用作重要的流行病学工具，用于检测患有腹泻住院的婴儿和幼儿的轮状病毒，并帮助确立了这些药物在婴儿和幼儿严重腹泻中的重要性。尽管已经做出了密集的努力，但在任何组织培养系统中尚未培养诺瓦克和相关的人类北病毒。这加上该群体某些成员的病毒样颗粒（VLP）的可视化，也有助于电子显微镜在盖子研究计划中持续重要的重要作用。电子显微镜在各种能力上一直是重要的主要和辅助工具。自去年的年度报告以来，研究包括各种领域：（1）在透析后进行纯化的制剂中，可视化完整的海象Vesivivirus VLP； （2）从2001年，2007年和2008年开始可视化澳大利亚彩色病毒VLP（与澳大利亚皇家儿童医院儿科的Mahers和Kirkwood女士合作）； （3）可视化Norwalk病毒壳和JHH3 P的嵌合VLP（与JHU的合作）； （4）试图确定抗坏血酸是否对轮状病毒SA-11在形态上有任何直接影响（与Niddk的Levine博士的合作）； （5）试图通过直接EM，通过IEM无离心的IEM和IEM进行离心（与Emerson博士的合作）来可视化丙型肝炎病毒； （6）试图确定丙型肝炎病毒基因型是否是IEM特有的基因型； （7）在杆状病毒系统中表达的Norwalk病毒 - 壳鼠Norovirus P结构域C嵌合膜状蛋白的可视化；（8）试图通过直接EM（与Lackdawala博士和Lamirande女士的协作）可视化流感病毒H1N1的形态学外观）； （9）在CSCL中的各种鼠病毒制剂的可视化；（10）在RML上为合作的冷冻研究准备了Norovirus JHH3 VLP制备的存在和完整性的可视化。