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Rapid and robust phylotyping of spa t003, a dominant MRSA clone in Luxembourg and other European countries.

ABSTRACT: spa typing is a common genotyping tool for methicillin-resistant Staphylococcus aureus (MRSA) in Europe. Given the high prevalence of dominant clones, spa-typing is proving to be limited in its ability to distinguish outbreak isolates from background isolates. New molecular tools need to be employed to improve subtyping of dominant local MRSA strains (e.g., spa type t003).Phylogenetically critical, or canonical, SNPs (can-SNPs) were identified as subtyping targets through sequence analysis of 40 MRSA whole genomes from Luxembourg. Real-time PCR assays were designed around target SNPs and validated using a repository of 240 previously sub-typed and epidemiologically characterized Luxembourg MRSA isolates, including 153 community and hospital isolates, 69 isolates from long term care (LTC) facilities, and 21 prospectively analyzed MRSA isolates. Selected isolates were also analyzed by whole genome SNP typing (WGST) for comparison to the SNP assays and other subtyping techniques.Fourteen real-time PCR assays were developed and validated, including two assays to determine presence of spa t003 or t008. The other twelve assays successfully provided a high degree of resolution within the t003 subtype. WGST analysis of the LTC facility isolates provided greater resolution than other subtyping tools, identifying clusters indicative of ongoing transmission within LTC facilities.canSNP-based PCR assays are useful for local level MRSA phylotyping, especially in the presence of one or more dominant clones. The assays designed here can be easily adapted for investigating t003 MRSA strains in other regions in Western Europe. WGST provides substantially better resolution than other typing methods.

SUBMITTER: Engelthaler DM

PROVIDER: S-EPMC3733620 | biostudies-literature | 2013 Jul

REPOSITORIES: biostudies-literature

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Rapid and robust phylotyping of spa t003, a dominant MRSA clone in Luxembourg and other European countries.

Engelthaler David M DM Kelley Erin E Driebe Elizabeth M EM Bowers Jolene J Eberhard Carl F CF Trujillo Jesse J Decruyenaere Frederic F Schupp James M JM Mossong Joel J Keim Paul P Even Jos J

BMC infectious diseases 20130723

<h4>Background</h4>spa typing is a common genotyping tool for methicillin-resistant Staphylococcus aureus (MRSA) in Europe. Given the high prevalence of dominant clones, spa-typing is proving to be limited in its ability to distinguish outbreak isolates from background isolates. New molecular tools need to be employed to improve subtyping of dominant local MRSA strains (e.g., spa type t003).<h4>Methods</h4>Phylogenetically critical, or canonical, SNPs (can-SNPs) were identified as subtyping targ ...[more]

PMID: 23879266

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Project description:BACKGROUND:The European Health Examination Survey in Luxembourg (EHES-LUX) is a population based survey performed from 2013 to 2015 with the aim to establish baseline information on the general health status of the Luxembourg population aged 25 to 64 years. The paper introduces the study design, recruitment method and representativeness of the sample, and summarizes the sociodemographic characteristics of participants and the prevalence of major health outcomes. METHODS:EHES-LUX is based on a random sample representative of the population of Luxembourg based on gender, age and district of residence. The sample size of the study was determined to provide accurate prevalence estimates for major chronic conditions. During two visits, data were collected from participants through a questionnaire (personal data, health status and health care), medical examinations (anthropometric measures, electrocardiogram and spirometry) and biological analysis (blood, urine and hair). Means and frequencies were used to describe the general characteristics of the population and a one-way ANOVA to test the representativeness of the sample and the comparability of participants and non-participants. RESULTS:A total of 1529 individuals participated in this study (participation rate of 24.1%). Differences between participants and non-participants based on gender, age and district of residence were corrected by sampling weights. The mean age (±SD) of participants was 44.9 (±10.1) years, of which 52.8% were women. Based on clinical measurements, nearly 20% of participants were obese and more than one in three participants were overweight. From measurements (respectively from self-report), 22.0% (respectively 12.2%) were hypertensive, 49.3% (respectively 22.5%) had hypercholesterolemia, 3.5% (respectively 3.7%) had diabetes and 7.6% (respectively 6.0%) had depressive symptoms. CONCLUSIONS:This nationwide epidemiological study on the general health status of Luxembourg residents provides updated prevalence estimates on a range of major health conditions. This information can be used by health authorities to evaluate policies and public health initiatives. At European level, prevalence data obtained by this study following the EHES-Reference Committee (EHES-RC) recommendations, will be comparable between European countries participating in this program.

Project description:Multiresistant Shiga toxin-producing Escherichia coli (STEC) O118:H16 and O118 nonmotile strains (designated O118:[H16]) were detected by examination of 171 STEC isolates for their antimicrobial sensitivity. Of 48 STEC O118:[H16] strains, 98% were resistant to sulfonamide, 96% were resistant to streptomycin, 79% were resistant to kanamycin, 75% were resistant to tetracycline, 67% were resistant to ampicillin, 60% were resistant to chloramphenicol, 48% were resistant to trimethoprim, and 10% each were resistant to gentamicin and nalidixic acid. Nalidixic acid resistance and reduced susceptibility to ciprofloxacin were associated with the mutation gyrA(LEU-83). The STEC O118:[H16] strains were found to belong to a single genetic clone as investigated by multilocus enzyme electrophoresis and by multilocus sequence analysis of E. coli housekeeping genes. The STEC O118:[H16] strains originated from humans and cattle and were isolated in seven different countries of Europe between 1986 and 1999. Strains showing multiresistance to up to eight different antimicrobials predominated among the more recent STEC O118:[H16] strains. The genes in parentheses were associated with resistance to kanamycin (aphA1-Ia), chloramphenicol (catA1), tetracycline [tet(A)], and ampicillin (bla(TEM-1)). Class 1 integrons containing sulI (sulfonamide resistance), aadA1a (streptomycin resistance), or dfrA1 (trimethoprim resistance)-aadA1a gene cassettes were detected in 28 strains. The bla(TEM-1b) gene was present in 18 of 21 strains that were examined by nucleotide sequencing. Class 1 integrons and bla(TEM) genes were localized on plasmids and/or on the chromosome in different STEC O118:[H16] strains. Hybridization of XbaI-digested chromosomal DNA separated by pulsed-field gel electrophoresis revealed that bla(TEM) genes were integrated at different positions in the chromosome of STEC O118:[H16] strains that could have occurred by Tn2 insertion. Our data suggest that strains belonging to the STEC O118:[H16] clonal group have a characteristic propensity for acquisition and maintenance of resistance determinants, thus contrasting to STEC belonging to other serotypes.

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Rapid and robust phylotyping of spa t003, a dominant MRSA clone in Luxembourg and other European countries.

Publications

Rapid and robust phylotyping of spa t003, a dominant MRSA clone in Luxembourg and other European countries.

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