Use of pyrosequencing to identify point mutations in domain V of 23S rRNA genes of linezolid-resistant Staphylococcus aureus and Staphylococcus epidermidis

A pyrosequencing assay was used for the rapid characterization of linezolid-resistant isolates of Staphylococcus aureus and Staphylococcus epidermidis. The assay identified base substitutions in copies of the 23S rRNA gene and determined the percentage of alleles with the mutation. Modifications of the assay were necessary to identify all mutations in the 23S rRNA genes of S. epidermidis that were associated with linezolid resistance. A C2534T mutation was identified in S. epidermidis that was not previously reported in a linezolid-resistant isolate.