Development and application of pathovar-specific monoclonal antibodies that recognize the lipopolysaccharide O antigen and the type IV fimbriae of <i>Xanthomonas hyacinthi</i>

作     者：van Doorn, J Ojanen-Reuhs, T Hollinger, TC Reuhs, BL Schots, A Boonekamp, PM Oudega, B 

作者机构：Bulb Res Ctr Dept Plant Qual NL-2160 AB Lisse Netherlands Univ Helsinki Dept Biosci Div Gen Microbiol SF-00014 Helsinki Finland Univ Georgia Complex Carbohydrate Res Ctr Athens GA 30602 USA Landbouw Univ Wageningen Dept Nematol NL-6700 ES Wageningen Netherlands Vrije Univ Amsterdam IMBW BCA Fac Biol NL-1081 HV Amsterdam Netherlands 

出 版 物：《APPLIED AND ENVIRONMENTAL MICROBIOLOGY》 (应用与环境微生物学)

年 卷 期：1999年第65卷第9期

页      面：4171-4180页

核心收录：

学科分类：0710[理学-生物学] 1007[医学-药学(可授医学、理学学位)] 100705[医学-微生物与生化药学] 07[理学] 0836[工学-生物工程] 071005[理学-微生物学] 10[医学] 

主　　题：抗体 细菌/免疫学 抗体 单克隆/免疫学 抗体特异性 抗原 细菌/分析 抗原 细菌/免疫学 酶联免疫吸附测定 表位/分析 表位/免疫学 纤毛 细菌/化学 纤毛 细菌/免疫学 免疫印迹法 磁共振波谱学 显微镜检查 免疫电子 O抗原/分析 O抗原/化学 O抗原/免疫学 植物疾病/微生物学 植物叶/微生物学 黄单胞菌属/免疫学 动物 小鼠 

摘      要：The objective of this study was to develop a specific immunological diagnostic assay for yellow disease in hyacinths, using monoclonal antibodies (MAbs). Mice were immunized with a crude cell wall preparation (shear fraction) from Xanthomonas hyacinthi and with purified type IV fimbriae. Hybridomas were screened for a positive reaction with X. hyacinthi cells or fimbriae and for a negative reaction with X. translucens pv. graminis or Erwinia carotovora subsp. carotovora. Nine MAbs recognized fimbrial epitopes, as shown by immunoblotting, immunofluorescence, enzyme-linked immunosorbent assay (ELISA), and immunoelectron microscopy;however, three of these MAbs had weak cross-reactions with two X. translucens pathovars in immunoblotting experiments. Seven MAbs reacted with lipopolysaccharides and yielded a low-mobility ladder pattern on immunoblots. Subsequent analysis of MAb 2E5 showed that it specifically recognized an epitope on the O antigen, which was found to consist of rhamnose and fucose in a 2:1 molar ratio. The cross-reaction of MAb 2E5 with all X. hyacinthi strains tested showed that this O antigen is highly conserved within this species. MAb 1B10 also reacted with lipopolysaccharides. MAbs 2E5 and 1B10 were further tested in ELISA and immunoblotting experiments with cells and extracts from other pathogens. No cross-reaction was found with 27 other Xanthomonas pathovars tested or with 14 other bacterial species from other genera, such as Erwinia and Pseudomonas, indicating the high specificity of these antibodies. MAbs 2E5 and 1B10 were shown to be useful in ELISA for the detection of X. hyacinthi in infected hyacinths.