Investigation of an outbreak of vancomycin-resistant Enterococcus faecium by random amplified polymorphic DNA (RAPD) assay

Issack, Mohammad Iqbal; Power, E.G.M.; French, Gary

doi:10.1016/s0195-6701(96)90002-x

Cited by 24 publications

(15 citation statements)

References 20 publications

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“…To investigate how MLST performs with epidemiologically related isolates, we analyzed 16 additional isolates (not belonging to the sample of 123 isolates) involved in different outbreaks in the United States, The Netherlands, Australia, and the United Kingdom and belonging to STs 16,17,18, and 20.…”

Section: T G C T C a C G T G G C C G C A C T G C G C T A A A A C Tmentioning

confidence: 99%

“…Of these, pulsed-field gel electrophoresis analysis of genomic restriction fragments has been considered the "gold standard" for the study of hospital outbreaks because of its high degree of isolate differentiation (15,17,20,23). However, due to this high degree of isolate differentiation, pulsed-field gel electrophoresis typing is less suitable for determining the degree of relatedness among epidemiologically unrelated isolates.…”

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confidence: 99%

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Multilocus Sequence Typing Scheme for Enterococcus faecium

Homan

Tribe²,

Poznanski³

et al. 2002

J Clin Microbiol

478

300

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A multilocus sequence typing (MLST) scheme has been developed for Enterococcus faecium. Internal fragments from seven housekeeping genes of 123 epidemiologically unlinked isolates from humans and livestock and 16 human-derived isolates from several outbreaks in the United States, the United Kingdom, Australia, and The Netherlands were analyzed. A total of 62 sequence types were detected in vancomycin-sensitive E. faecium (VSEF) and vancomycin-resistant E. faecium (VREF) isolates. VSEF isolates were genetically more diverse than VREF isolates. Both VSEF and VREF isolates clustered in host-specific lineages that were similar to the host-specific clustering obtained by amplified fragment length polymorphism analysis. Outbreak isolates from hospitalized humans clustered in a subgroup that was defined by the presence of a unique allele from the housekeeping gene purK and the surface protein gene esp. The MLST results suggest that epidemic lineages of E. faecium emerged recently worldwide, while genetic variation in both VREF and VSEF was created by longer-term recombination. The results show that MLST of E. faecium provides an excellent tool for isolate characterization and long-term epidemiologic analysis.Vancomycin-resistant Enterococcus faecium (VREF) has recently emerged as an important threat in U.S. hospitals (5, 24). In Europe, VREF isolates are found relatively frequently in the community and farm animals, while prevalence in hospitals is generally low (14). The latter observation was explained by the use of the glycopeptide avoparcin as an antimicrobial growth promoter in animal feeding operations.Several molecular typing schemes have been developed to study the epidemiology of VREF. Of these, pulsed-field gel electrophoresis analysis of genomic restriction fragments has been considered the "gold standard" for the study of hospital outbreaks because of its high degree of isolate differentiation (15,17,20,23). However, due to this high degree of isolate differentiation, pulsed-field gel electrophoresis typing is less suitable for determining the degree of relatedness among epidemiologically unrelated isolates. Recently, amplified fragment length polymorphism (AFLP) analysis was applied as a new method for the typing of VREF (1, 33). AFLP analysis is a robust and fast typing technique with high intra-and interexperimental reproducibilities and appears to be discriminatory enough for the recognition of hospital outbreaks (1, 32, 33). In addition, AFLP analysis has allowed the detection of associations among different E. faecium genetic lineages and different human and animal hosts (33), suggesting the existence of host-specific VREF lineages. Whether this is also true for vancomycin-sensitive E. faecium (VSEF) is not known, since VSEF isolates were not included in that study. AFLP typing also disclosed two different human-associated lineages. One lineage comprised epidemic-related isolates recovered from hospitalized patients, while isolates of the other lineage were mainly from nonhospitalized persons. Interes...

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Section: T G C T C a C G T G G C C G C A C T G C G C T A A A A C Tmentioning

confidence: 99%

mentioning

confidence: 99%

Multilocus Sequence Typing Scheme for Enterococcus faecium

Homan

Tribe²,

Poznanski³

et al. 2002

J Clin Microbiol

478

300

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show abstract

“…Amplified fragment length polymorphism (AFLP) is a typing method that utilizes a combination of restriction enzyme digestion and PCR (21,42,56,137,196,270,287,309,315,317). In the AFLP procedure, the DNA is digested with two different restriction endonucleases, usually chosen so that one cuts more frequently than the other.…”

Section: Typing Methods Using Pcrmentioning

confidence: 99%

Application of Molecular Techniques to the Study of Hospital Infection

et al. 2006

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SUMMARY Nosocomial infections are an important source of morbidity and mortality in hospital settings, afflicting an estimated 2 million patients in United States each year. This number represents up to 5% of hospitalized patients and results in an estimated 88,000 deaths and 4.5 billion dollars in excess health care costs. Increasingly, hospital-acquired infections with multidrug-resistant pathogens represent a major problem in patients. Understanding pathogen relatedness is essential for determining the epidemiology of nosocomial infections and aiding in the design of rational pathogen control methods. The role of pathogen typing is to determine whether epidemiologically related isolates are also genetically related. To determine molecular relatedness of isolates for epidemiologic investigation, new technologies based on DNA, or molecular analysis, are methods of choice. These DNA-based molecular methodologies include pulsed-field gel electrophoresis (PFGE), PCR-based typing methods, and multilocus sequence analysis. Establishing clonality of pathogens can aid in the identification of the source (environmental or personnel) of organisms, distinguish infectious from noninfectious strains, and distinguish relapse from reinfection. The integration of molecular typing with conventional hospital epidemiologic surveillance has been proven to be cost-effective due to the associated reduction in the number of nosocomial infections. Cost-effectiveness is maximized through the collaboration of the laboratory, through epidemiologic typing, and the infection control department during epidemiologic investigations.

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“…(PyMS), multi-locus enzyme electrophoresis, analysis of whole or digested plasmid DNA, restriction endonuclease In one of the earliest studies of enterococcal transmission Gross et al (1976) showed that four of nine patients with analysis of chromosomal DNA, ribotyping, IS6770 probing, digestion of amplified fragments of glycopeptide resistance indwelling urinary catheters had contamination of the perineum, meatus and parts of the catheter. As many as 10% of genes, various pulsed-field gel electrophoresis techniques (PFGE) and PCR typing (Woodford et al 1995;Issack et al physicians and nursing personnel carried enterococci on their hands. However, with one exception, they all were different 1996; Morrison et al 1996b).…”

Section: Typing Methodsmentioning

confidence: 99%