Clonal nature and diversity of resistance, toxins and adhesins genes of meticillin-resistant Staphylococcus aureus collected in a Spanish hospital

doi:10.1016/j.meegid.2012.07.020

Infection, Genetics and Evolution

Volume 12, Issue 8, December 2012, Pages 1751-1758

https://doi.org/10.1016/j.meegid.2012.07.020 Get rights and content

Abstract

In this study we determined the prevalence of genes coding for antimicrobial resistance, toxins, enzymes, immunoevasion and adhesins factors among 189 meticillin-resistant Staphylococcus aureus (MRSA) strains isolated from a third level hospital in Valladolid (Spain) between 2005 and 2008 in order to examine the relationship between these pathogenicity determinants, both individually and in combination, and the genetic background of main MRSA strains that are presents in Spanish hospitals. MRSA isolates were first characterised epidemiologically by a combination of molecular typing strategies like spa, SCCmec and multilocus sequence typing, and then, a cluster analysis based on pathogenicity factors genes was performed according to the hybridisation pattern of 65 virulence, 36 resistance, 15 adhesins, and 11 set/ssl genes on a Diagnostic DNA microarray (Alere StaphyType DNA microarray Jena, Germany). CC5-agr type II [ST125-SCCmecIV/VI (32.2%) or ST125-IV (19.1%), ST228-I (19.1%), ST146-IV (13.7%) and ST5- IV (0.5%)] isolates was widely distributed. CC8-agr type I [ST8-IV (11.5%), USA300 clone (0.5%), and ST239-III (1.1%)]; CC45-agr type II [ST45- IV (1.6%)], and the CC97-agr type I [ST97-IV] were also detected. We identified 42 different resistance genes profiles, 22 set/ssl genes profiles, and 91 different virulence profiles. However although the high genetic diversity of MRSA strains, mainly with respect to virulence factors genes, the results of the simultaneous assessment of resistance and virulence genes and the genetic background illustrated a correspondence relationship (p < 0.001) between the different clones and same resistance and virulence genes or clusters of them. During the study period we observed changes in molecular epidemiology of MRSA isolates and as a consequence we report the changes of the resistance and virulence potential of MRSA strains produced over time in our institution.

Highlights

► We determine the prevalence of pathogenicity genes among 189 MRSA isolates. ► 91 Virulence, 42 resistance and 22 set/ssl genes profiles were detected. ► We observed a correspondence relationship between clones and pathogenicity genes. ► Changes in molecular epidemiology in our institution were produced over time. ► We reported the pathogenic potential of prevalent MRSA clones in Spanish hospitals.

Introduction

Methicillin-resistant Staphylococcus aureus (MRSA) has become established as one of the most prevalent pathogen in hospitals worldwide. Aspects related to these infections have important epidemiologic, therapeutic and clinical implications, both from the perspective of its pathogenicity as well as from the evolution of antimicrobial resistance. In the past two decades, MRSA strains have generally been confined to healthcare settings (HA-MRSA) and have predominantly affected individuals with co-morbidity or other specific risk factors, such as prolonged hospital stay and nursing home residency (Elston and Barlow, 2009). However, over the years, MRSA infection epidemiology has undergone important changes. Novel strains have also emerged outside of the hospital setting, in the community (CA-MRSA) (David and Daum, 2010), and more recently livestock animals have been described as a novel source of human MRSA colonization and infection (LA-MRSA) (Graveland et al., 2011). CA-MRSA and LA-MRSA have become a challenge for countries that have so far maintained low rates of MRSA (Stefani et al., 2012). Also, CA-MRSA strains are gradually becoming entrenched as nosocomial pathogens and that both CA-MRSA and HA-MRSA strains now circulate in both environments (David et al., 2008).

The evolution of new human and animal pathogenic strains of S. aureus has been due to the accumulation of mobile genetic elements encoding methicillin resistance and virulence factors into successful lineages (Lindsay, 2010). Also, recent analyses have shown that all S. aureus genotypes that efficiently colonize humans have given rise to life-threatening pathogens, but that some clonal lineages appear to be more virulent than others (Holtfreter et al., 2007). Comparative genomics has been an increasingly important approach for scientists to improve knowledge on the pathogenesis and drug resistance of S. aureus.

The aim of this study was to characterise both resistance and virulence genetic profiles of MRSA clones isolated in a third level hospital in Valladolid, Spain between 2005 and 2008, in order to obtain an insight into the presence and genetic characteristics of epidemic MRSA strains in Spain

Section snippets

Bacterial isolates

A total of 189 non repeated MRSA isolates recovered from clinical samples were collected in 2005, 2007 and 2008 at the University Clinic Hospital in Valladolid, Spain (HCUV). Isolates were considered to be nosocomially acquired in 126 cases, healthcare-associated in 47 cases, community-associated in two cases, and of uncertain mode of acquisition in fourteen. The isolates were reported to originate from skin and soft tissue infection (105 isolates), surgery infection (48), respiratory tract

Molecular typing

spa Typing of the 189 MRSA strain studied revealed 18 different spa types (Table 1). Representative MRSA isolates analyzed by MLST showed eight different sequence types (ST125, ST5, ST146, ST228, ST8, ST239, ST45 and ST97) which belonged to four clonal complexes (CC5, CC8, CC45 and CC97). The strains studied were classified as belonging to nine different HA-MRSA clones, designated by their ST followed by their SCCmec type (ST125-IV/VI, ST125-IV, ST146-IV, ST228-I, ST5-IV, ST8-IV, ST239-III,

Discussion

The vast majority of our MRSA isolates could be attributed to five clones ST125-IV/VI, ST125-IV, ST228-I, ST146-IV and ST8-IV. The ST125 isolates are currently more frequent in our hospital, as well as in other Spanish hospitals, which represents almost the 50% of MRSA isolates (Perez-Vazquez et al., 2009, Vindel et al., 2009). However, unlike that observed in other Spanish hospitals, we find that 62% of these ST125 strains harbored composite or multiple SCCmec elements including SCCmec type IV

Acknowledgements

This study was partially supported by a Grant of Roche Diagnostic.

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S.aureusST97 has been described as associated with bovine mastitis in many parts of the world (Hata et al., 2010, for review see Cuny et al., 2009). More recently it was also reported from pigs in Germany (Meemken et al., 2010) and in Spain where it has been found among isolates from nasal carriers (Lozano et al., 2011) and with low frequency also among isolates from nosocomial infections (Menegotto et al., 2012). In the United States MRSA ST97 has been reported as a contaminant from urban environmental surfaces of a student's campus (Roberts et al., 2011) and from babies in a neonatal unit in Kuwait (Udo et al., 2011).
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Clonal nature and diversity of resistance, toxins and adhesins genes of meticillin-resistant Staphylococcus aureus collected in a Spanish hospital

Abstract

Highlights

Introduction

Section snippets

Bacterial isolates

Molecular typing

Discussion

Acknowledgements

J. Mol. Biol.

Blood

J. Infect.

Int. J. Med. Microbiol.

Int. J. Med. Microbiol.

Clin. Microbiol. Infect.

Clin. Microbiol. Infect.

Clin. Microbiol. Infect.

Methods Enzymol.

Int. J. Antimicrob. Agents

The interaction between staphylococcal superantigen-like proteins and human dendritic cells

Clin. Exp. Immunol.

The crystal structure of staphylococcal superantigen-like protein 11 in complex with Sialyl Lewis X reveals the mechanism for cell binding and immune inhibition

Mol. Microbiol.

Community-associated methicillin-resistant Staphylococcus aureus: epidemiology and clinical consequences of an emerging epidemic

Clin. Microbiol. Rev.

What is community-associated methicillin-resistant Staphylococcus aureus?

J. Infect. Dis.

Nucleotide sequence of the Staphylococcus aureus transposon, Tn5405, carrying aminoglycosides resistance genes

J. Basic. Microbiol.