Studies Of Gastroenteritis Viruses By Electron Microscopy

通过电子显微镜研究胃肠炎病毒

• 批准号: 8156828
• 负责人: Albert Z Kapikian
• 金额: $ 19.74万
• 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/8156828
• 关键词: Electron Microscopy; Gastroenteritis; Virus

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 expression of 27nm 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. In last year's annual report, we cited the various uses of the electron microscope for different studies including examination of (1) VLP preparations prepared from Children's Hospital archival GII-4 specimens; (2) a high-titered tetravalent human-bovine rotavirus (UK) reassortant vaccine preparation from a licensee to determine if any contaminant could be seen, to assist in an investigation of the cause of guinea pig toxicology issues; (3) VLP preparations of various caliciviruses (Norwalk virus, MD 145 virus, mink calicivirus, murine norovirus and others); (4) stool specimen from a chimpanzee with non A-E hepatitis by IEM with a pre and two post infection sera in an attempt to detect a viral agent (in collaboration with Dr. Purcell); (5) a virus seed of a primary African Green monkey kidney grown human-bovine reassortant rotavirus in an investigation of an adventitious agent; (6) purified dengue 4 vaccine preparations (in collaboration with Dr. Anez Gutierrez); (7) hepatitis E virus preparation (in collaboration with Dr. Emerson). This year the use of the electron microscope has had a similar varied theme with examination of (1) putative VLP preparations of various caliciviruses (Norwalk virus, Toronto virus, archival specimens of GII-3 noroviruses from the 1980s and 1990s, Desert Shield for use in chimpanzee studies, Hom-1 virus, walrus vesivirus, Akita sapovirus); (2) D X porcine Gottfried reassortant vaccine candidate (Dr.Hoshino); (3) D x porcine Gottfried reassortant (ether-treated x 2) for screening of titer; (4) D x porcine Gottfried reassortant from terminal dilution study; (5) 2 archival anal swab suspensions derived from an outbreak of gastroenteritis in 1966 on the destroyer Shenandoah (this was the earliest gastroenteritis outbreak evaluated by LID in volunteers); (6) stool suspensions derived from a tertiary case in a gastroenteritis outbreak involving children and adults by direct EM initially and later by IEM; (7) mink calicivirus; (8) hepatitis E virus in an IEM study (in collaboration with Dr. Emerson (MHS).

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。这以及该组剂的某些成员的27nm病毒样颗粒（VLP）的表达促成了电子显微镜在盖子研究计划中的持续重要作用。电子显微镜在各种能力上一直是重要的主要和辅助工具。在去年的年度报告中，我们引用了电子显微镜在不同研究中的各种用途，包括对（1）VLP的研究。 （2）从被许可人准备的高素质四位型人类轮状病毒（英国）重新排育疫苗的准备，以确定是否可以看到任何污染物，以协助调查豚鼠毒理学问题； （3）各种孔孔病毒的VLP制剂（Norwalk病毒，MD 145病毒，Mink Calicivirus，Murine Norovirus等）； （4）来自IEM的黑猩猩的粪便标本，伴有IEM的非A-E肝炎，并在感染前和两个后感染血清试图检测病毒剂（与Purcell博士合作）； （5）一种非洲绿猴肾脏的病毒种子在调查一个不定代理的人类种植的人群轮状病毒中； （6）纯化的登革热4疫苗制剂（与Anez Gutierrez博士合作）； （7）乙型肝炎病毒的制备（与艾默生博士合作）。 今年，电子显微镜的使用具有类似的主题，并检查了（1）各种孔孔病毒（Norwalk病毒，多伦多病毒，多伦多病毒，1980年代和1990年代的诺维病毒的档案标本）的假定VLP制剂，沙漠盾牌用于黑猩猩研究，hom-hom-1 （2）D X猪Gottfried候选疫苗（Dr.Hoshino）； （3）D X猪Gottfried Reastortant（经以太处理的X 2）用于筛选滴度； （4）D X猪Gottfried从末端稀释研究中恢复； （5）2档案肛门拭子悬浮液在1966年在驱逐舰Shenandoah爆发的胃炎爆发（这是LID在志愿者中评估的最早的胃肠炎爆发）； （6）在胃炎爆发中涉及第三病例的粪便悬浮液最初和以后的IEM涉及儿童和成人； （7）貂皮炎病毒； （8）在IEM研究中（与Emerson博士（MHS）合作，乙型肝炎病毒。