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A number of Enterococcus strains with high-level inducible resistance to vancomycin have been identified, and the relative incidence of these strains has increased significantly in the last years. The first outbreak caused by vancomycin-resistant enterococci in Poland was reported in 1999. Vancomycin-resistant Enterococcus faecium is known for its propensity to cause infections which are difficult to eradicate. In this study, we determined the genetic similarities between vancomycin-resistant E. faecium isolates consecutively recovered from single patients to assess the duration of infection or colonization. The isolates taken in the study were identified by the conventional methods as E. faecium. PCRmelting profile (PCR-MP) and pulsed-field gel electrophoresis (PFGE) typing revealed that the isolates belonged to six distinct genotypes and that two of them were predominant. Consecutive E. faecium isolates with identical genotypes were found in 7 of 12 (58.0%) patients. The delay between the times of recovery of the first and last isolates of identical genotypes from each patient was from 9 days to about 1 year. In six patients, paired blood and non-blood isolates showed identical genotypes. Data presented here demonstrate the complexity of the epidemiological situation concerning vancomycin-resistant enterococci that may occur in a single medical ward. We also show for the first time the evaluation of PCR-MP technique in enterococci strains differentiation and we revealed that there is at least a similar power of discrimination between the presentjjold-standard REA-PFGE and a PCR-MP method.
The development of rapid and simple typing methods is required in order to identify possible sources of human exposure to opportunistic pathogens. Klebsiella spp. belongs to a group of bacteria that are opportunistic pathogens responsible for an increasing number of multi-resistant infections in hospitals. Recently, we showed the high genetic diversity of K. oxytoca using a large collection of strains isolated from the patients of several hospitals in Poland over a 50-year period. Our results showed that the internal transcribed spacer polymerase chain reaction method (ITS-PCR) is useful for the phylogenetic delineation of genetic groups in K. oxytoca and the high discriminatory power of the PCR melting profiles (PCR MP) method can be useful for epidemiological studies of K. oxytoca. In the present study the usefulness of PCR MP was tested on two sets of strains isolated from a single unit over a short period of time. The results revealed that PCR MP has a high discriminatory power and can be useful for epidemiological studies of closely related strains of K. oxytoca isolated from a single unit over a short period of time to identify the source, reservoirs and the tract of infection spread. The advantage of PCR MP for the above application was shown by using the procedure at increasing denaturation temperature during PCR to confirm genotyping results. Considering this feature and the high discriminatory power of PCR MP, as shown in this report for determination of the genetic similarities of consecutive K. oxytoca strains, we propose that PCR MP is one of the best techniques for short-term epidemiology analysis.
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