Kuipers B, van den Dobbelsteen G, Wedege E, van Alphen L
Laboratory of Vaccine Research, National Institute of Public Health and the Environment, Bilthoven, The Netherlands, Oslo, Norway.
Methods Mol Med. 2001;67:131-45. doi: 10.1385/1-59259-149-3:131.
Immunoassays employ a range of methods to detect and quantify antigens or antibodies and to study the composition of antigens. This chapter describes four useful immunoassays for serological characterization of antigens of Neisseria meningitidis: whole-cell enzyme-linked immunosorbent assay (ELISA); dot blot, colony blot, and immunoblot. Serological characterization of N. meningitidis antigens is valuable for epidemiological studies as well as for identifying immunologically important antigens in vaccine development (1,2). Typing of N. meningitidis is based on the immunological detection of specific epitopes on the outer-membrane proteins (OMP) and lipopolysaccharides (LPS) (3), and panels of monoclonal antibodies (MAbs) have been developed by several laboratories to refine the serological classification system (4-8). Differences in capsular polysaccharides determine the meningococcal serogroup, whereas the serotypes and serosubtypes are based on antigenic differences of the PorB OMP and PorA OMP, respectively (3). The PorA protein contains two variable loops (VR1 and VR2), each of which determine a dis- tinct set of serosubtypes. Thus the serosubtypes of an isolate can include two independent designations (9-11). Variation in the oligosaccharide moiety of the LPS determines the immunotype, and more than one epitope can be present in the same population of a single isolate (6,12). In the current typing scheme the classification is given as [serogroup]: [serotype]: [serosubtype]: [immunotype], e.g., B: 15:P1.7,16:L3,7,9.
免疫测定采用一系列方法来检测和定量抗原或抗体，并研究抗原的组成。本章描述了四种用于脑膜炎奈瑟菌抗原血清学特征分析的有用免疫测定方法：全细胞酶联免疫吸附测定（ELISA）；斑点印迹、菌落印迹和免疫印迹。脑膜炎奈瑟菌抗原的血清学特征分析对于流行病学研究以及在疫苗开发中鉴定具有免疫学重要性的抗原都很有价值(1,2)。脑膜炎奈瑟菌的分型基于对外膜蛋白（OMP）和脂多糖（LPS）上特定表位的免疫检测(3)，几个实验室已经开发了单克隆抗体（MAb）组来完善血清学分类系统(4 - 8)。荚膜多糖的差异决定了脑膜炎球菌的血清群，而血清型和血清亚型分别基于PorB OMP和PorA OMP的抗原差异(3)。PorA蛋白包含两个可变环（VR1和VR2），每个可变环决定一组不同的血清亚型。因此，分离株的血清亚型可以包括两个独立的命名(9 - 11)。LPS寡糖部分的变异决定了免疫型，并且在单个分离株的同一群体中可以存在不止一个表位(6,12)。在当前的分型方案中，分类表示为[血清群]：[血清型]：[血清亚型]：[免疫型]，例如，B：15：P1.7,16：L3,7,9。
