Characterisation of MRSA strains isolated from patients in a hospital in Riyadh, Kingdom of Saudi Arabia.

BACKGROUND: Methicillin-resistant Staphylococcus aureus (MRSA) is spreading worldwide and poses a serious public health problem, being present in hospital settings and communities. However, from the Middle East and the Arabian Peninsula few molecular typing data on MRSA strains are currently available. In order to obtain data on the population structure of MRSA in Riyadh, Saudi Arabia, 107 clinical and environmental MRSA isolates were genotyped using a microarray-based assay. RESULTS: Five major MRSA strains from four clonal complexes were identified CC8/ST239-III (20.75%), PVL-positive as well as -negative CC22-IV (18.87% and 9.43%, respectively), PVL-positive CC30-IV (12.26%) and PVL-positive CC80-IV (17.92%). Minor strains, which accounted for less than 3% each, included CC1-IV/SCCfus, PVL-positive CC1/ST772-V, PVL-positive as well as- negative CC5-IV, CC5-IV/SCCfus, CC5-V, CC6-IV, CC45-IV, PVL-negative CC80-IV, PVL-positive CC88-IV, CC97-V and a CC9/ST834-MRSA strain. CONCLUSIONS: Typing of MRSA strains from Riyadh revealed a high diversity of clonal complexes. The prevalence of the genes encoding the Panton-Valentine leukocidin was surprisingly high (54.21%), and a significant rate of resistance markers was detected also in strains considered as community-associated.

Characterization of PVL-positive MRSA from Norway.

Norway is a country in which the Methicillin-resistant Staphylococcus aureus (MRSA) prevalence has been low for the last decades. There are virtually no epidemic, hospital-acquired MRSA because of an emphasis on strict infection control rules and restrictive use of antibiotics. However, community-acquired and/or Panton-Valentine leucocidin (PVL)-positive MRSA need to be monitored as these strains are transmitted outside of healthcare facilities and cannot be contained by healthcare-centred strategies. All 179 non-repetitive isolates of PVL-positive MRSA that were received during 2011 at the regional infection control laboratory at Akershus University Hospital were preserved and spa typed. Seventy isolates were further characterized by DNA microarray hybridization. The most common PVL-MRSA lineages were ST8-MRSA-IV and CC30-MRSA-IV. Further common clones were CC80-MRSA-IV and CC5-MRSA-IV. Other clones were found sporadically. These included ST772-MRSA-V and ST834-MRSA-IV, the latter in patients with epidemiological connections to the Philippines. Small-scale family outbreaks affecting at least 49 individuals were noted, with numbers of known cases per outbreak ranging from two to seven. At least 24 cases were related to foreign travel to Eritrea, India, Iraq, Macedonia, Pakistan, the Philippines, Poland, Singapore, Turkey, the USA and Vietnam. These data show that community-acquired/PVL-positive MRSA are not yet a major public health problem in Southern Norway. Our study corroborates the current practice of mandatory screening of patients and staff with travel histories, admissions or employment in healthcare institutions outside the Scandinavian countries or with known MRSA contacts.

Rapid identification of carbapenemase genes in gram-negative bacteria with an oligonucleotide microarray-based assay.

Rapid molecular identification of carbapenemase genes in Gram-negative bacteria is crucial for infection control and prevention, surveillance and for epidemiological purposes. Furthermore, it may have a significant impact upon determining the appropriate initial treatment and greatly benefit for critically ill patients. A novel oligonucleotide microarray-based assay was developed to simultaneously detect genes encoding clinically important carbapenemases as well as selected extended (ESBL) and narrow spectrum (NSBL) beta-lactamases directly from clonal culture material within few hours. Additionally, a panel of species specific markers was included to identify Escherichia coli, Pseudomonas aeruginosa, Citrobacter freundii/braakii, Klebsiella pneumoniae and Acinetobacter baumannii. The assay was tested using a panel of 117 isolates collected from urinary, blood and stool samples. For these isolates, phenotypic identifications and susceptibility tests were available. An independent detection of carbapenemase, ESBL and NSBL genes was carried out by various external reference laboratories using PCR methods. In direct comparison, the microarray correctly identified 98.2% of the covered carbapenemase genes. This included blaVIM (13 out of 13), blaGIM (2/2), blaKPC (27/27), blaNDM (5/5), blaIMP-2/4/7/8/13/14/15/16/31 (10/10), blaOXA-23 (12/13), blaOXA-40-group (7/7), blaOXA-48-group (32/33), blaOXA-51 (1/1) and blaOXA-58 (1/1). Furthermore, the test correctly identified additional beta-lactamases [blaOXA-1 (16/16), blaOXA-2 (4/4), blaOXA-9 (33/33), OXA-10 (3/3), blaOXA-51 (25/25), blaOXA-58 (2/2), CTX-M1/M15 (17/17) and blaVIM (1/1)]. In direct comparison to phenotypical identification obtained by VITEK or MALDI-TOF systems, 114 of 117 (97.4%) isolates, including Acinetobacter baumannii (28/28), Enterobacter spec. (5/5), Escherichia coli (4/4), Klebsiella pneumoniae (62/63), Klebsiella oxytoca (0/2), Pseudomonas aeruginosa (12/12), Citrobacter freundii (1/1) and Citrobacter braakii (2/2), were correctly identified by a panel of species specific probes. This assay might be easily extended, adapted and transferred to point of care platforms enabling fast surveillance, rapid detection and appropriate early treatment of infections caused by multiresistant Gram-negative bacteria.

Genotyping of Staphylococcus aureus isolates from diseased poultry.

To gain insight into the genomic diversity of Staphylococcus aureus associated with diseases in domestic poultry, 131 isolates from clinically ill turkeys (n=80) and chickens (n = 51) were collected and genotyped using microarray hybridisations. MRSA isolates were subjected to spa and dru typing and their antimicrobial resistance geno- and phenotypes were determined. Most (68 out of 80) turkey isolates belonged to the clonal complex (CC) 398. Seventeen of the 80 isolates (21.2%) were MRSA. The most common MRSA type among turkeys was CC398-MRSA-V (n = 8), but CC5-MRSA-III (n = 4), CC9-MRSA-IV (n = 2), CC398-MRSA-IV (n = 2) and a single CC398-MRSA with an unidentified/truncated SCCmec element were also found. Among the chicken isolates, CC5 predominated (44 out of 51). Five of the chicken isolates were MRSA (9.8%), all belonging to CC398-MRSA-V. These data show that the current dissemination of livestock-associated MRSA also engulfs chickens and turkeys, and that MRSA surveillance among these species is warranted.

A field guide to pandemic, epidemic and sporadic clones of methicillin-resistant Staphylococcus aureus.

In recent years, methicillin-resistant Staphylococcus aureus (MRSA) have become a truly global challenge. In addition to the long-known healthcare-associated clones, novel strains have also emerged outside of the hospital settings, in the community as well as in livestock. The emergence and spread of virulent clones expressing Panton-Valentine leukocidin (PVL) is an additional cause for concern. In order to provide an overview of pandemic, epidemic and sporadic strains, more than 3,000 clinical and veterinary isolates of MRSA mainly from Germany, the United Kingdom, Ireland, France, Malta, Abu Dhabi, Hong Kong, Australia, Trinidad & Tobago as well as some reference strains from the United States have been genotyped by DNA microarray analysis. This technique allowed the assignment of the MRSA isolates to 34 distinct lineages which can be clearly defined based on non-mobile genes. The results were in accordance with data from multilocus sequence typing. More than 100 different strains were distinguished based on affiliation to these lineages, SCCmec type and the presence or absence of PVL. These strains are described here mainly with regard to clinically relevant antimicrobial resistance- and virulence-associated markers, but also in relation to epidemiology and geographic distribution. The findings of the study show a high level of biodiversity among MRSA, especially among strains harbouring SCCmec IV and V elements. The data also indicate a high rate of genetic recombination in MRSA involving SCC elements, bacteriophages or other mobile genetic elements and large-scale chromosomal replacements.