Westprinting: Development of a rapid immunochemical identification for species within the genus Pseudomonas sensu stricto

A strategy determined Westprinting (for Westernblot and immunological fingerprinting) has been proposed for the immunological identification of the authentic genus Pseudomonas (rRNA-DNA homology group I; Proteobacteria gamma-3 subclass) over a wide phylogenetic range. For that purpose an antiserum has been raised which generated genus- and species-specific protein profiles of all members of the phylogenetic group after sodium dodecyl-sulfate polyacrylamide-gel electrophoresis (SDS-PAGE) and immunoblotting. Cluster analyses based on Westprints of different Pseudomonas type strains revealed similar interspecies and intra-species relationships comparable to 16S rRNA gene sequence analysis. Differentiation of several strains belonging to the same species was shown for P. fluorescens, as well as for seven genomovars of P. stutzeri which have been well characterized by DNA-DNA similarity analysis and 16S rRNA gene-sequencing. The method has also been used for a rapid and reliable screening assay and multiple environmental isolates were identified to the genus and species level. On the basis of the Westprinting technology Pseudomonas- characteristic antigens were identified and monoclonal antibodies were raised against outer membrane protein (Opr) H2 and a 11 kDa protein band, which reacted against all examined pseudomonads belonging to rRNA-DNA homology group I.