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PCR melting profile method for genotyping analysis of vancomycin-resistant Enterococcus faecium isolates from
Beata Krawczyk1, Justyna Leibner, Karolina Stojowska
1Gdańsk University of Technology, Department of Microbiology, Gdańsk, Poland.
Abstract:
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. PCR melting 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 present gold-standard REA-PFGE and a PCR-MP method.
Insights
High-level vancomycin-resistant Enterococcus faecium can persist in patients for up to a year, complicating infections. Genetic analysis showed identical strains can cause prolonged colonization or infection in over half of patients studied.
Area of Science:
- Microbiology
- Infectious Diseases
- Epidemiology
Background:
- Increasing incidence of vancomycin-resistant enterococci (VRE), particularly Enterococcus faecium, poses significant challenges in infection control.
- VRE infections are notoriously difficult to eradicate, necessitating better understanding of their persistence.
- The first VRE outbreak in Poland was reported in 1999, highlighting the growing public health concern.
Purpose of the Study:
- To determine the genetic relatedness of vancomycin-resistant E. faecium isolates from individual patients.
- To assess the duration of infection or colonization by genetically similar VRE strains.
- To evaluate the discriminatory power of PCR melting profile (PCR-MP) compared to pulsed-field gel electrophoresis (PFGE).
Main Methods:
- Conventional identification of Enterococcus faecium isolates.
- Genotyping using PCR melting profile (PCR-MP) and pulsed-field gel electrophoresis (PFGE).
- Analysis of consecutively recovered isolates from 12 patients to assess strain persistence and genetic similarity.
Main Results:
- Six distinct genotypes of vancomycin-resistant E. faecium were identified, with two predominant genotypes.
- Identical genotypes were found in 58% of patients, with persistence ranging from 9 days to approximately one year.
- Paired blood and non-blood isolates from six patients shared identical genotypes.
- PCR-MP demonstrated a discriminatory power comparable to the gold-standard REA-PFGE for enterococci strain differentiation.
Conclusions:
- Vancomycin-resistant E. faecium can persist within a single patient for extended periods, contributing to complex epidemiological situations in healthcare settings.
- The PCR-MP technique is a reliable and effective method for differentiating enterococci strains, offering an alternative to PFGE.
- Understanding the duration of VRE colonization/infection is crucial for effective infection control strategies.
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