Detection of novel chromosome-SCCmec variants in Methicillin Resistant Staphylococcus aureus and their inclusion in PCR based screening

Findings. To facilitate automation, a novel DNA extraction method for MRSA was adopted. The MRSA specific chromosome-SCCmec PCR was adapted, additional primers were added, and the performance was validated. From various laboratories in The Netherlands we received a total of 86 MRSA clinical isolates, that were negative in commercially available tests. We identified 14 MRSA strains with new variant chromosome-SCCmec junctions by sequence analysis. These MRSA strains appeared to carry SCCmec sequences with a high degree of homology to SCC regions of S. epidermidis and S. haemolyticus. All were included for detection in chromosome-SCCmec based PCR. Background: Efficient management of Methicillin Resistant Staphylococcus aureus (MRSA) in the hospital is needed to prevent dissemination. It is important that MRSA can be rapidly identified, and effective infection control measures can be initiated. Equally important is a rapid MRSA negative report, especially for patients in isolation. For negative screening we implemented fully automated high through-put molecular screening for MRSA. Conclusions: Fourteen variant chromosome-SCCmec junctions in MRSA, that are not detected in commercially available MRSA detection kits were added to our PCR to detect all currently known variant SCC-mec types of MRSA

Whole-genome sequencing of bacterial pathogens:the future of nosocomial outbreak analysis

SUMMARY Outbreaks of multidrug-resistant bacteria present a frequent threat to vulnerable patient populations in hospitals around the world. Intensive care unit (ICU) patients are particularly susceptible to nosocomial infections due to indwelling devices such as intravascular catheters, drains, and intratracheal tubes for mechanical ventilation. The increased vulnerability of infected ICU patients demonstrates the importance of effective outbreak management protocols to be in place. Understanding the transmission of pathogens via genotyping methods is an important tool for outbreak management. Recently, whole-genome sequencing (WGS) of pathogens has become more accessible and affordable as a tool for genotyping. Analysis of the entire pathogen genome via WGS could provide unprecedented resolution in discriminating even highly related lineages of bacteria and revolutionize outbreak analysis in hospitals. Nevertheless, clinicians have long been hesitant to implement WGS in outbreak analyses due to the expensive and cumbersome nature of early sequencing platforms. Recent improvements in sequencing technologies and analysis tools have rapidly increased the output and analysis speed as well as reduced the overall costs of WGS. In this review, we assess the feasibility of WGS technologies and bioinformatics analysis tools for nosocomial outbreak analyses and provide a comparison to conventional outbreak analysis workflows. Moreover, we review advantages and limitations of sequencing technologies and analysis tools and present a real-world example of the implementation of WGS for antimicrobial resistance analysis. We aimed to provide health care professionals with a guide to WGS outbreak analysis that highlights its benefits for hospitals and assists in the transition from conventional to WGS-based outbreak analysis. </jats:sec

Clonal Distribution and Differential Occurrence of the Enterotoxin Gene Cluster, egc, in Carriage- versus Bacteremia-Associated Isolates of Staphylococcus aureus

The Staphylococcus aureus enterotoxin gene cluster, egc, was detected in isolates from healthy individuals and in those from patients with bacteremia. The egc genes cooccur and are slightly enriched in strains from healthy carriers (present in 63.7% of carriage-associated isolates versus 52.9% of invasive isolates; P = 0.03). Multilocus sequence typing revealed that successful staphylococcal clones usually harbor the egc locus

Nasopharyngeal co-colonization with Staphylococcus aureus and Streptococcus pneumoniae in children is bacterial genotype independent.

Item does not contain fulltextBacterial interference between Staphylococcus aureus and Streptococcus pneumoniae in the nasopharynx has been observed during colonization, which might have important clinical implications for the widespread use of pneumococcal conjugate vaccine in young children. This study aimed to determine whether the capacity of Staph. aureus to compete with Strep. pneumoniae is dependent on bacterial genotype. Demographic and microbiological determinants of carriage of specific genotypes of Staph. aureus in children were also studied. Children (n=3198) were sampled in the nasopharynx to detect carriage of Staph. aureus, Strep. pneumoniae and Neisseria meningitidis. Staph. aureus genotypes and pneumococcal sero- and genotypes were determined. Age, gender, zip code, active smoking and co-colonization with N. meningitidis or Strep. pneumoniae, both vaccine- and non-vaccine types, were not associated with colonization by specific Staph. aureus genotypes. Based on the whole-genome typing data obtained, there was no obvious correlation between staphylococcal and pneumococcal genotypes during co-colonization. Passive smoking showed a significant association (P=0.003) with carriage of a specific Staph. aureus cluster. This study suggests that there are no major differences between Staph. aureus clones (with different disease-invoking potential) in their capacity to compete with Strep. pneumoniae subtypes. Further studies should demonstrate whether differences in bacterial interference are due to more subtle genetic changes