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Antimicrobial Agents and Chemotherapy, February 2005, p. 536-540, Vol. 49, No. 2
0066-4804/05/$08.00+0     doi:10.1128/AAC.49.2.536-540.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Mutations in folP Associated with Elevated Sulfonamide MICs for Neisseria meningitidis Clinical Isolates from Five Continents

Department of Pathology, The University of Texas Health Science Center, San Antonio, Texas

Received 16 August 2004/ Returned for modification 3 October 2004/ Accepted 17 October 2004

Sulfonamide resistance in meningococci is associated with mutations in the chromosomal gene folP, which encodes dihydropteroate synthase. Several mutations associated with resistance have been previously described, including amino acid substitutions at codons 31 and 194, a glycine-serine insertion at codons 195 and 196, and, recently, an additional mutation at nucleotide 682 (C682A). In this study, sulfisoxazole MICs were determined for 424 geographically diverse clinical isolates of Neisseria meningitidis, including all major subtypes. A subset of 134 isolates with MICs ranging from 0.5 to >64 µg/ml were assayed for the C682A mutation by real-time PCR, and 25 isolates were selected for folP gene sequencing. All isolates for which the sulfisoxazole MIC was 8 possessed the C682A mutation by real-time PCR or folP sequencing, and 34 of 35 isolates with a MIC of 2 lacked this mutation. Of 16 sequenced isolates for which the sulfisoxazole MIC was 4, 15 possessed previously described mutations, including 10 at codon 31, 1 at codon 194, and 4 with the 2-amino-acid insertion codons 195 and 196; all 16 possessed the C682A mutation. The C682A mutation predicted elevated sulfonamides MICs for a large number of geographically diverse clinical isolates of meningococci. Detection of this mutation by real-time PCR or other methods may allow more wide-scale detection of meningococcal isolates with for which the sulfonamide MICs are elevated without resorting to multiple assays or folP gene sequencing, providing a simple, high-throughput screening method for use in public health and epidemiologic settings.

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