Li Z, Jensen N S, Bélanger M, L'Espérance M C, Jacques M
Groupe de Recherche sur les Maladies Infectieuses du Porc, Faculté de Médecine Vétérinaire, Université de Montréal, Saint-Hyacinthe, Québec.
J Clin Microbiol. 1992 Nov;30(11):2941-7. doi: 10.1128/jcm.30.11.2941-2947.1992.
The study described here was carried out to further characterize reference strains of Serpulina (Treponema) hyodysenteriae representing serotypes 8 and 9. Results obtained from restriction fragment length polymorphism analysis, enteropathogenicity testing, and endotoxin profiles confirmed their identifications. Electron microscopy indicated that both strains were covered with a thin layer of capsule-like material. Immunoblot analysis indicated that an antigen in the 19-kDa region of proteinase K-digested whole cells reacted only with homologous antiserum. The serotype-specific antigens were sensitive to periodate oxidation but resistant to proteinase K digestion and migrated in the same region as purified lipopolysaccharides. Immunoblotting with proteinase K-digested whole cells appeared as useful as immunodiffusion with extracted lipopolysaccharide for the serological classification of S. hyodysenteriae. Immunogold labeling of whole cells and purified periplasmic flagella showed strong cross-reactions between S. hyodysenteriae and Serpulina innocens. Outer membrane preparations of strains representing serotypes 8 and 9 contained four major proteins which reacted with antisera against both species, and one major protein with a molecular mass of 46 kDa which reacted only with antisera against S. hyodysenteriae, irrespective of the serotype. Our findings suggest that periplasmic flagella and some outer membrane proteins are antigens common to both S. hyodysenteriae and S. innocens, whereas a 46-kDa outer membrane protein may be a species-specific antigen of S. hyodysenteriae. Finally, we propose immunoblotting as an alternative method to immunodiffusion for the serotyping of S. hyodysenteriae.
本研究旨在进一步鉴定猪痢疾蛇形螺旋体（密螺旋体属）8型和9型参考菌株的特征。通过限制性片段长度多态性分析、肠致病性检测和内毒素谱分析所获得的结果证实了它们的特性。电子显微镜检查表明，两株菌均覆盖有一层薄的荚膜样物质。免疫印迹分析表明，蛋白酶K消化的全细胞19 kDa区域内的一种抗原仅与同源抗血清发生反应。血清型特异性抗原对高碘酸盐氧化敏感，但对蛋白酶K消化有抗性，且与纯化的脂多糖在同一区域迁移。对于猪痢疾蛇形螺旋体的血清学分类，蛋白酶K消化的全细胞免疫印迹与提取的脂多糖免疫扩散一样有用。全细胞和纯化的周质鞭毛的免疫金标记显示猪痢疾蛇形螺旋体和无害蛇形螺旋体之间有强烈的交叉反应。8型和9型菌株的外膜制剂含有四种主要蛋白质，它们与两种菌的抗血清都发生反应，还有一种分子量为46 kDa的主要蛋白质，它仅与猪痢疾蛇形螺旋体的抗血清发生反应，而与血清型无关。我们的研究结果表明，周质鞭毛和一些外膜蛋白是猪痢疾蛇形螺旋体和无害蛇形螺旋体共有的抗原，而一种46 kDa的外膜蛋白可能是猪痢疾蛇形螺旋体的种特异性抗原。最后，我们建议将免疫印迹作为猪痢疾蛇形螺旋体血清分型的一种替代免疫扩散的方法。
