Green K Y, Lew J F, Jiang X, Kapikian A Z, Estes M K
Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, Bethesda, Maryland 20892.
J Clin Microbiol. 1993 Aug;31(8):2185-91. doi: 10.1128/jcm.31.8.2185-2191.1993.
Since the discovery of the Norwalk virus (NV) by immune electron microscopy (IEM) in 1972, serologic studies with this virus have relied on particle-positive fecal material from infected volunteers as the source of antigen because it has not been possible to propagate this virus in cell culture. However, the recent cloning of the NV (strain 8FIIa) genome and expression of the capsid protein in a baculovirus system to form "virus-like particles" has provided a consistent source of antigen (designated rNV). The purpose of the present study was to compare the antigenicities of these rNV particles with those of native NV antigen derived from human fecal material by using well-characterized sera obtained from earlier studies. In IEM studies, the rNV antigen reacted with NV-specific antibodies in a manner similar to that observed previously when particle-positive fecal material was used as antigen. In addition, a direct enzyme-linked immunosorbent assay, in which the rNV antigen was used as antigen, proved efficient and specific for the detection of serologic responses to NV compared with the previously established techniques of IEM and blocking antibody immunoassays in which particle-positive fecal material was used as the antigen. The availability of an unlimited source of antigen will enable serologic studies that will greatly increase our understanding of the epidemiology of NV and its role in human enteric illness.
自1972年通过免疫电子显微镜(IEM)发现诺沃克病毒(NV)以来,针对该病毒的血清学研究一直依赖于感染志愿者的粪便样本作为抗原来源,因为这种病毒无法在细胞培养中繁殖。然而,最近NV(8FIIa株)基因组的克隆以及衣壳蛋白在杆状病毒系统中的表达,形成了“病毒样颗粒”,提供了一种稳定的抗原来源(称为rNV)。本研究的目的是通过使用早期研究中获得的特征明确的血清,比较这些rNV颗粒与源自人类粪便的天然NV抗原的抗原性。在IEM研究中,rNV抗原与NV特异性抗体的反应方式,与之前使用颗粒阳性粪便样本作为抗原时观察到的方式相似。此外,与之前使用颗粒阳性粪便样本作为抗原的IEM和阻断抗体免疫测定技术相比,以rNV抗原作为抗原的直接酶联免疫吸附测定法,在检测针对NV的血清学反应方面,被证明是有效且特异的。无限量抗原来源的可用性,将使血清学研究得以开展,这将极大地增进我们对NV流行病学及其在人类肠道疾病中作用的理解。