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Antimicrobial Agents and Chemotherapy, November 2007, p. 3969-3976, Vol. 51, No. 11
0066-4804/07/$08.00+0     doi:10.1128/AAC.00422-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Genetic Characteristics and Clonal Dissemination of ß-Lactamase-Negative Ampicillin-Resistant Haemophilus influenzae Strains Isolated from the Upper Respiratory Tract of Patients in Japan

Department of Otolaryngology-Head and Neck Surgery, Wakayama Medical University, Wakayama, Japan,¹ Surveillance Subcommittee, Japan Society for Infectious Diseases in Otolaryngology,² Department of Otolaryngology, Second Affiliated Hospital, Fujita Health University, Nagoya, Japan³

Received 28 March 2007/ Returned for modification 30 April 2007/ Accepted 27 July 2007

We evaluated the recent prevalence of antimicrobial-resistant Haemophilus influenzae isolated from the upper respiratory tracts (URT) of patients in Japan. Mutations in the ftsI gene, which encodes penicillin binding protein 3 (PBP3), and the clonal dissemination of the resistant strains were also investigated. A total of 264 H. influenzae isolates were collected from patients with URT infections. According to the criteria of the Clinical and Laboratory Standards Institute for the susceptibility of H. influenzae to ampicillin (AMP), the isolates were distributed as follows: 161 (61.0%) susceptible strains (MIC 1 µg/ml), 37 (14.0%) intermediately resistant strains (MIC = 2 µg/ml), and 66 (25.0%) resistant strains (MIC 4 µg/ml). According to PCR-based genotyping, 172 (65.1%) of the isolates had mutations in the ftsI gene and were negative for the ß-lactamase (bla) gene. These 172 isolates were thus defined as genetically ß-lactamase-negative ampicillin-resistant (gBLNAR) strains. The ftsI mutant group included 98 (37.1%) strains with group I/II mutations in the variable mutated region (group I/II gBLNAR) and 74 (28.0%) strains with group III mutations in the highly mutated region (group III gBLNAR). Eighty-seven (33.0%) of the isolates were genetically ß-lactamase-negative ampicillin-susceptible (gBLNAS) strains. The group III gBLNAR strains showed resistance to ß-lactams. Only five strains (1.9%) were positive for a bla gene encoding TEM-type ß-lactamase. The three clusters consisting of 16 strains found among the 61 BLNAR strains (MIC 4 µg/ml and without the bla gene) showed identical or closely related DNA restriction fragment patterns. Those isolates were frequently identified among strains with a MIC to AMP of 16 µg/ml. The current study demonstrates the apparent dissemination and spread of a resistant clone of H. influenzae among medical centers in Japan. The gBLNAR strains show a remarkable prevalence among H. influenzae isolates, with the prevalence increasing with time. This fact should be taken into account when treating URT infections.

Published ahead of print on 13 August 2007.

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