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Multiplex PCR Assay for Typing of Staphylococcal Cassette Chromosome Mec Types I to V in Methicillin-resistant Staphylococcus aureus
Published on: September 5, 2013
Systematic derivation of marker sets for staphylococcal cassette chromosome mec typing
Alex J Stephens1, Flavia Huygens, Philip M Giffard
1Institute of Health and Biomedical Innovation, Queensland University of Technology, Kelvin Grove, QLD, Australia.
Abstract:
The aim of this study was to identify optimized sets of genotyping targets for the staphylococcal cassette chromosome mec (SCCmec). We analyzed the gene contents of 46 SCCmec variants in order to identify minimal subsets of targets that provide useful resolution. This was achieved by firstly identifying and characterizing each available SCCmec element based on the presence or absence of 34 binary targets. This information was used as input for the software "Minimum SNPs," which identifies the minimum number of targets required to differentiate a set of genotypes up to a predefined Simpson's index of diversity (D) value. It was determined that 22 of the 34 targets were required to genotype the 46 SCCmec variants to a D of 1. The first 6, 9, 12, and 15 targets were found to define 21, 29, 35, and 39 SCCmec variants, respectively. The genotypes defined by these marker subsets were largely consistent with the relationships between SCCmec variants and the accepted nomenclature. Consistency was made virtually complete by forcing the computer program to include ccr1 and ccr5 in the target set. An alternative target set biased towards discriminating abundant SCCmec variants was derived by analyzing an input file in which common SCCmec variants were repeated, thus ensuring that markers that discriminate abundant variants had a large effect on D. Finally, it was determined that mecA single nucleotide polymorphisms (SNPs) can increase the overall genotyping resolution, as different mecA alleles were found in otherwise identical SCCmec variants.
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