Evaluation of different PCR-based approaches for the identification and typing of environmental enterococci.
The aim of the work was the evaluation of different PCR-based methods to found an appropriate identification and typing strategy for environmental enterococci. Environmental enterococci were isolated mainly from surface- and waste-waters. Species identification was provided by combination of phenotypic (Micronaut System, Merlin) and molecular detection methods (fluorescent ITS-PCR, ddl-PCR, REP-PCR, AFLP). Very similar results were observed among molecular methods, however several discrepancies were recognized during comparison of molecular and biochemical identification. Seven enterococcal species (E. faecium, E. hirae, E. casseliflavus, E. mundtii, E. faecalis, E. durans and E. gallinarum) were identified within 166 environmental isolates. The results obtained in this work attest the importance of PCR-based methods for identification and typing of environmental enterococci. The fluorescent ITS-PCR (fITS-PCR) showed the best results in order to identify the enterococci strains, the method used the automated capillary electrophoresis to separate the PCR products in a very rapid and precise way. The AFLP method was suitable to identify and characterize the isolates, while the REP-PCR can be used for species identification.
Pangallo
Domenico
D
Institute of Molecular Biology, Slovak Academy of Sciences, Dúbravská cesta 21, Bratislava 84551, Slovak Republic. domenico.pangallo@savba.sk
Drahovska
Hana
H
Harichova
Jana
J
Karelova
Edita
E
Chovanova
Katarina
K
Aradska
Jana
J
Ferianc
Peter
P
Turna
Jan
J
Timko
Jozef
J
D021903:DNA, Ribosomal Spacer / Q000737:chemistry / Q000235:genetics; D016983:Enterococcus / Q000145:classification / Q000235:genetics*; D005838:Genotype; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA
D021903:DNA, Ribosomal Spacer