Abstract

BackgroundStaphylococcal protein A (spa) is an important virulence factor which enables Staphylococcus aureus to evade host immune responses. Genotypes known as “spa-types”, based on highly variable Xr region sequences of the spa-gene, are frequently used to classify strains. A weakness of current spa-typing primers is that rearrangements in the IgG-binding region of the gene cause 1-2% of strains to be designated as “non-typeable”.ResultsWe developed an improved primer which enabled sequencing of all strains, containing any type of genetic rearrangement, in a large study among community carriers and hospital inpatients in Oxfordshire, UK (6110 isolates). We identified eight novel spa-gene variants, plus one previously described. Three of these rearrangements would be designated “non-typeable” using current spa-typing methods; they occurred in 1.8% (72/3905) asymptomatically carried and 0.6% (14/2205) inpatient S. aureus strains. Some individuals were simultaneously colonized by both formerly non-typeable and typeable strains; previously such patients would have been identified as carrying only currently typeable strains, underestimating mixed carriage prevalence and diversity. Formerly non-typeable strains were found in more spa-types associated with multilocus sequence type ST398 (35%), common among livestock, compared to other groups with any non-typeable strains (1-4%), suggesting particular spa-types may have been under-represented in previous human studies.ConclusionsThis improved method allows us to spa-type previously non-typeable strains with rearrangements in the spa-gene and to resolve cases of mixed colonization with deletions in one or more strains, thus accounting for hidden diversity of S. aureus in both community and hospital environments.

Highlights

  • Staphylococcal protein A is an important virulence factor which enables Staphylococcus aureus to evade host immune responses

  • Deletions appear to be over-represented in clonal lineages related to livestock In total, we found 20 spa-types from 33 individuals associated with nine types of rearrangements in the spa-gene (Additional file 2: Table S2)

  • Spa-typing of 6110 S. aureus isolates showed that 1.8% of samples from 1.8% community carriers and 0.6% of samples from 0.7% inpatients were formerly non-typeable

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Summary

Introduction

Staphylococcal protein A (spa) is an important virulence factor which enables Staphylococcus aureus to evade host immune responses. A weakness of current spa-typing primers is that rearrangements in the IgG-binding region of the gene cause 1-2% of strains to be designated as “non-typeable”. S. aureus encodes many virulence factors including the surface Ig-binding protein A (spa) whose function is to capture IgG molecules in the inverted orientation and Typing the highly variable Xr region of the spa-gene is one of the most common methods for genotyping S. aureus. One weakness of current spa-typing methods is that rearrangements in the in the IgG-binding region of the gene, where the forward spa-primer is located, lead to. Five nonspa-typeable S. aureus clinical strains with rearrangements in the IgG-binding domain of the spa-gene were first described by Baum et al in 2009 [14]. Whilst the prevalence of such rearrangements can be directly estimated from the proportion of non-typeable strains, detecting rearrangements that do not affect spa-typing would require sequencing the whole spa-gene; such rearrangements may still be informative with respect to population structure

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