Sugiyama T, Kido N, Arakawa Y, Mori M, Naito S, Ohta M, Kato N
Department of Bacteriology, Nagoya University School of Medicine.
Microbiol Immunol. 1990;34(7):635-41. doi: 10.1111/j.1348-0421.1990.tb01039.x.
A rapid small-scale method of extraction of lipopolysaccharide (LPS) and capsular polysaccharides was developed for the purpose of identification of chemotypes of LPS and serotypes of capsular antigens. Cell surface polysaccharides were prepared within less than 2 hr from 1.5 ml of broth or suspension of colonies cultured overnight. The preparations were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis for LPS, and by double diffusion gel precipitation (Ouchterlony) test and blotting to nitrocellulose membrane for capsular polysaccharide. The analyses with the preparations obtained by the method could provide adequate results capable of identifying chemotypes of LPS and serotypes of capsular antigens.
为了鉴定脂多糖(LPS)的化学型和荚膜抗原的血清型,开发了一种快速小规模提取脂多糖和荚膜多糖的方法。从1.5 ml过夜培养的肉汤或菌落悬液中,在不到2小时内制备细胞表面多糖。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析脂多糖,通过双向扩散凝胶沉淀(Ouchterlony)试验和硝酸纤维素膜印迹分析荚膜多糖。用该方法获得的制剂进行分析能够提供足以鉴定脂多糖化学型和荚膜抗原血清型的结果。