ABSTRACT:The purpose of this study was to evaluate the discriminatory power of ddl-PCR (D-alanin-D-alanin ligase PCR) and ITS-PCR (Internal Transcribed Spacer PCR) for accurate identification of enterococcal species in comparison with phenotypic assays. Results confirm previous published data that ddl-PCR simple approach allows rapid identification of two the most frequently isolated spp. E. faecalis and E. faecium. When identification points towards other enterococci then that mentioned above, the ITS-PCR seems to be suitable complementary assay. Correct identification of enterococci to species level is important because of different spp. susceptibility to some clinically important antibiotics. For example, it is necessary to distinquish acquired vancomycin resistance from inherent one that is less epidemiologically important. Additionally, both methods can be valuable in epidemiological studies following enterococcus resistance gene transfer within human population.

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The genes coding for 4 aminoglycoside-modifying enzymes AAC(6')-APH(2"), APH(3'), ANT(4') and ANT(6) were determined in 44 Slovak clinical isolates of Enterococcus faecalis with high-level resistance to gentamicin (HLGR, collection 1) and 48 E. faecalis isolates with resistance to amikacin (AR, collection 2). The occurrence of spotted genes was (collection 1 vs. collection 2): aac(6)-aph(2") 81.8 vs. 8.3 %, ant(4') 52.3 vs. 81.3 %, aph(3') 50 vs. 56.3 % and ant(6) 6.8 vs. 4.2 %, the most frequent combinations of genes in the HLGR collection were aac(6')-aph(2") + ant(4') and aac(6')-aph(2") + aph(3). In contrast, the aph(3') + ant(4') gene profile was predominant in AR isolates. None of the isolates contained all four AGME genes simultaneously.

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