Yoda T, Terano Y, Suzuki Y, Yamazaki K, Oishi I, Utagawa E, Shimada A, Matsuura S, Nakajima M, Shibata T
Division of Food Microbiology, Osaka Prefectural Institute of Public Health, Japan.
Microbiol Immunol. 2000;44(11):905-14. doi: 10.1111/j.1348-0421.2000.tb02582.x.
The Norwalk virus (NV) causes outbreaks of acute non-bacterial gastroenteritis in humans. The virus capsid is composed of a single 60 kDa protein. The capsid protein of NV36 (genogroup II, Mexico virus type) was expressed in an Escherichia coli system and ten monoclonal antibodies (MAbs) were generated against it. The reactivity of these MAbs was characterized using enzyme-linked immunosorbent assay (ELISA) and Western blot (WB) analysis towards 20 overlapping fragments of the NV36 capsid protein expressed in E. coli. All of the MAbs recognized sequential (continuous) epitopes on the three antigenic regions. Six of the 10 MAbs recognized fragment 2 (equivalent residues 31-70), three MAbs recognized fragment 13 (residues 361-403) and one MAb recognized fragment 7 (residues 181-220), suggesting that the N-terminal domain (residues 1-220) may contain more antigenic epitopes than the C-terminal domain (residues 210-548). Furthermore, two MAbs (1B4 and 1F6) reacted in WB with three purified NV strains (genogroup II) derived from patients' stool samples. It was also found that genogroup I recombinant NV96-908 (genogroup I, KY89 type) could be detected as sensitively as recombinant NV36 (genogroup II) by ELISA with a set of the MAbs produced here.
诺如病毒（NV）可引发人类急性非细菌性肠胃炎疫情。该病毒衣壳由单一的60 kDa蛋白组成。NV36（基因群II，墨西哥病毒型）的衣壳蛋白在大肠杆菌系统中表达，并由此产生了十种单克隆抗体（MAb）。利用酶联免疫吸附测定（ELISA）和蛋白质印迹（WB）分析法，针对在大肠杆菌中表达的NV36衣壳蛋白的20个重叠片段，对这些单克隆抗体的反应性进行了表征。所有单克隆抗体均识别三个抗原区域上的连续（线性）表位。10种单克隆抗体中有6种识别片段2（对应残基31 - 70），3种单克隆抗体识别片段13（残基361 - 403），1种单克隆抗体识别片段7（残基181 - 220），这表明N端结构域（残基1 - 220）可能比C端结构域（残基210 - 548）含有更多的抗原表位。此外，两种单克隆抗体（1B4和1F6）在蛋白质印迹中与源自患者粪便样本的三种纯化NV毒株（基因群II）发生反应。还发现，利用此处产生的一组单克隆抗体，通过ELISA检测基因群I重组NV96 - 908（基因群I，KY89型）时，其灵敏度与重组NV36（基因群II）相同。
