Multilocus enzyme analysis in aerobic and anaerobic bacteria using gel electrophoresis-nitrocellulose blotting

An optimized multilocus enzyme electrophoresis method, which involves polya crylamide-agarose gel electrophoresis followed by electrophoretic transfers on nitrocellulose sheets, was developed for the analysis of enzyme polymor phism in several aerobic and anaerobic bacterial species including Staphylo coccus aureus, Streptococcus pneumoniae, S. agalactiae, Klebsiella pneumoni ae and K. oxytoca, Clostridium bifermentans and C. sordellii, and Prevotell a bivia. Serial electrophoretic transfers (during 5-15 min each) from a sin gle polyacrylamide gel could be achieved for most enzymes studied, and allo wed an increased definition of enzyme bands on nitrocellulose as compared t o migration gels. Four enzymes, which could not be blotted in such conditio ns, could still be stained in gels after blotting. Thus, the method allowed the combined analysis of several enzymes after a single gel electrophoresi s separation. The analysis of enzyme polymorphism in the various species st udied raised the interest of polymorphic loci such as esterase or glutamic- oxaloacetic transaminase for epidemiologic studies. The method characterize d a genetic diversity of enzyme loci of S. pneumoniae higher than previousl y reported, and is thus convenient for the analysis of genetic relationship s between related isolates. Since the present method reduces the tediousnes s of multilocus enzyme electrophoresis and requires experimental conditions that are not specific for the bacterial population studied, it may be prop osed for rapid population genetics analysis of a wide variety of bacteria. (C) 2000 Federation of European Microbiological Societies. Published by Els evier Science B.V. All rights reserved.