Pyle S W, Schill W B
J Immunol Methods. 1985 Dec 27;85(2):371-82. doi: 10.1016/0022-1759(85)90146-2.
Techniques are described for the rapid screening of proteinase K-treated bacterial lysates by electroblot and immunoenzymatic detection to assess O-specificity of antigens and antisera. Conditions are outlined which permit the use of a single polyacrylamide gel for both electrotransfer to nitrocellulose and silver staining. Immunodetection of transferred LPS bands was equally sensitive to silver stain when whole cell or O-specific antisera were used. The techniques were utilized to identify at least 4 O-serotypes among sorbitol fermenting isolates of the fish pathogen, Yersinia ruckeri. Observed variations in the electrophoretic mobilities of lipopolysaccharides from 17 field isolates of Y. ruckeri were used to accurately predict the O-serotype.
描述了通过电印迹和免疫酶检测快速筛选蛋白酶K处理的细菌裂解物以评估抗原和抗血清的O特异性的技术。概述了允许使用单一聚丙烯酰胺凝胶进行电转移至硝酸纤维素膜和银染色的条件。当使用全细胞或O特异性抗血清时,转移的脂多糖带的免疫检测对银染色同样敏感。这些技术被用于鉴定鱼类病原体鲁氏耶尔森菌的山梨醇发酵分离株中的至少4种O血清型。观察到的来自17株鲁氏耶尔森菌田间分离株的脂多糖电泳迁移率变化被用于准确预测O血清型。