S. aureus Genotyping Kit 2.0

(item number 245200096)

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.

This kit allows DNA-based detection of resistance genes and pathogenicity markers of Staphylococcus aureus and assignment of unknown S. aureus isolates to known strains. The set of target genes consists of a variety of species markers, virulence-associated genes including genes encoding PVL, exotoxins, antibiotic resistance genes, genes encoding microbial surface components recognizing adhesive matrix molecules (MSCRAMMs), as well as of other markers. DNA from S. aureus colony material is amplified and labelled, and resulting amplicons are hybridised to DNA oligonucleotide microarrays with 336 probes. Spot recognition is performed automatically based on digital images of the arrays. The overall pattern is analysed automatically for the presence or absence of specific genes and it is compared to a database of strain profiles allowing assignment to clonal complexes (as defined by MLST) and strains.

In the recent (2.0) version of the assay, additional probes for the newly described methicillin resistance gene mecC were introduced and the database was expanded accordingly. This gene that also confers resistance to beta lactam compounds in S. aureus was discovered in 2011 (Shore 2011, García-Álvarez 2011). It is localised on a novel SCCmec element designated as type XI. Because of its highly divergent sequence, it cannot be detected by routinely used molecular assays designed to identify mecA. This gene has been found S. aureus belonging to clonal complexes 49, 130, 425, 599 and 1943. MRSA that carry mecC have been isolated from humans as well as from various wild and domestic animals from Western Europe.

The assay can be used for research and for epidemiological / public health applications, for obtaining insight into S. aureus / MRSA population structures in defined settings or conditions as well as for the confirmation of the presence of certain virulence- or resistance-associated genes in clinical isolates.

A list of target genes is provided in the manual. The database of recognised clonal complexes and strains is expanded on a regular basis, and they are also listed in the manual.

Publications

Detection of staphylococcal cassette chromosome mec type XI encoding highly divergent mecA, mecI, mecR1, blaZ and ccr genes in human clinical clonal complex 130 methicillin-resistant Staphylococcus aureus
Shore AC, Deasy EC, Slickers P, Brennan G, O'Connell B, Monecke S, Ehricht R, Coleman DC. Antimicrobial Agents and Chemotherapy, published ahead of print on 2 June 2011, doi:10.1128/AAC.00187-11.

A field guide to pandemic, epidemic and sporadic clones of methicillin-resistant Staphylococcus aureus 
Monecke S, Coombs G, Shore AC, Coleman DC, Akpaka P, Borg M, Chow H, Ip M, Jatzwauk L, Jonas D, Kadlec K, Kearns A, Laurent F, O'Brien FG, Pearson J, Ruppelt A, Schwarz S, Scicluna E, Slickers P, Tan HL, Weber S, Ehricht R. PLoS One. 2011 Apr 6; 6(4):e17936. doi: 10.1371/journal.pone.0017936. PMID: 21494333

Clonal replacement of epidemic methicillin-resistant Staphylococcus aureus strains in a German university hospital over a period of eleven years 
Albrecht N, Jatzwauk L, Slickers P, Ehricht R, Monecke S.
PLoS One. 2011; 6(11):e28189. doi: 10.1371/journal.pone.0028189. Epub 2011 Nov 30. PMID: 22140542

Assignment of Staphylococcus aureus isolates to clonal complexes based on microarray analysis and pattern recognition 
Monecke S, Slickers P, Ehricht R. FEMS Immunol Med Microbiol. 2008 Jul; 53(2):237-51. doi: 10.1111/j.1574-695X.2008.00426.x. Epub 2008 May 27. PMID: 18507678

List of referring online publications.

Array-on-demand

Alere Technologies provides customised manufacturing of microarrays mounted in ArrayTube, ArrayTube2 and/or ArrayStrip. You decide which markers/genes you focus on. It is possible to adapt the existing assays/gene lists according to your requirements!

Genotyping Service

Please send either a clonal culture (e.g., suitable agar plate, stab culture or swab) or a high quality DNA preparation into our lab. We do the genotyping and you get a result file in electronic form. On request, we will perform a phenotypical characterization of antibiotic susceptibility and/or species identification by a VITEK-2 System (bioMérieux Deutschland GmbH).


For further questions please contact us!