Rapid serological analysis of bacterial lipopolysaccharides by electrotransfer to nitrocellulose

Abstract

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.