WESTPRINTING - DEVELOPMENT OF A RAPID IMMUNOCHEMICAL IDENTIFICATION FOR SPECIES WITHIN THE GENUS PSEUDOMONAS-SENSU-STRICTO

Citation
M. Tesar et al., WESTPRINTING - DEVELOPMENT OF A RAPID IMMUNOCHEMICAL IDENTIFICATION FOR SPECIES WITHIN THE GENUS PSEUDOMONAS-SENSU-STRICTO, Systematic and applied microbiology, 19(4), 1996, pp. 577-588
Citations number
64
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Microbiology,"Biothechnology & Applied Migrobiology
ISSN journal
0723-2020
Volume
19
Issue
4
Year of publication
1996
Pages
577 - 588
Database
ISI
SICI code
0723-2020(1996)19:4<577:W-DOAR>2.0.ZU;2-8
Abstract
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; Proteo bacteria gamma-3 subclass) over a wide phylogenetic range. For that pu rpose an antiserum has been raised which generated genus- and species- specific protein profiles of all members of the phylogenetic group aft er sodium dodecyl-sulfate polyacrylamide-gel electrophoresis (SDS-PAGE ) and immunoblotting. Cluster analyses based on Westprints of differen t Pseudomonas type strains revealed similar interspecies and intra-spe cies relationships comparable to 16S rRNA gene sequence analysis. Diff erentiation of several strains belonging to the same species was shown for P. fluorescens, as well as for seven genomovars of P. stutzeri wh ich have been well characterized by DNA-DNA similarity analysis and 16 S rRNA gene-sequencing. The method has also been used for a rapid and reliable screening assay and multiple environmental isolates were iden tified to the genus and species level. On the basis of the Westprintin g 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 pseu domonads belonging to rRNA-DNA homology group I.