The resistance mechanisms and clonal relationship were determined for two Streptococcus pyogenes isolates with high-level resistance to levofloxacin as well as to other fluoroquinolones. DNA amplification and sequencing revealed a serine-81 -> phenylalanine substitution in GyrA and a double substitution in ParC of serine-79 -> phenylalanine and aspartic acid-91 -> asparagine. Pulsed-field gel electrophoresis analysis and emm typing determined that both isolates were emm type 28 and were genetically indistinguishable. (c) 2007 Elsevier B.V. and the International Society of Chemotherapy. All rights reserved.