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Journal of Clinical Microbiology, March 2003, p. 1167-1172, Vol. 41, No. 3
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.3.1167-1172.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Identification of a Mutation Associated with Erythromycin Resistance in Bordetella pertussis: Implications for Surveillance of Antimicrobial Resistance

J. M. Bartkus,1* B. A. Juni,1 K. Ehresmann,1 C. A. Miller,1 G. N. Sanden,2 P. K. Cassiday,2 M. Saubolle,3 B. Lee,4 J. Long,5 A. R. Harrison, Jr.,6,1 and J. M. Besser1

Minnesota Department of Health, Minneapolis,1 Crossroads Medical Center, Chaska, Minnesota,6 Centers for Disease Control and Prevention,2 Children's Hospital, Atlanta, Georgia,5 Good Samaritan Medical Center, Phoenix, Arizona,3 Oakland Children's Hospital, Oakland, California4

Received 10 September 2002/ Returned for modification 25 October 2002/ Accepted 17 December 2002

Erythromycin treatment failures and in vitro resistance of Bordetella pertussis have been reported on several occasions in the past few years, but the mechanism of resistance has not been described. One potential mechanism, genetic modification of the erythromycin-binding site on the 23S rRNA of the 50S ribosomal subunit, has been observed in other bacteria. To explore this possibility, we amplified the portion of the 23S rRNA gene encoding the central loop of domain V. DNA sequencing and restriction fragment length polymorphism of the PCR products showed that each of the four erythromycin-resistant B. pertussis strains tested contained an A-to-G transition mutation at position 2058 (Escherichia coli numbering) of the 23S rRNA gene. The mutation was not found in seven erythromycin-susceptible isolates tested. Two of the resistant isolates were heterozygous, containing at least one mutant copy and one wild-type copy of the 23S rRNA gene. These results indicate that erythromycin resistance in these strains is likely due to a mutation of the erythromycin-binding site in the 23S rRNA gene. Identification of the resistance mechanism will facilitate development of molecular susceptibility testing methods that can be used directly on clinical specimens in the absence of an isolate.

* Corresponding author. Mailing address: Public Health Laboratory, Minnesota Department of Health, 717 Delaware St., S.E., P.O. Box 9441, Minneapolis, MN 55440. Phone: (612) 676-5249. Fax: (612) 676-5514. E-mail: joanne.bartkus{at}health.state.mn.us.

{dagger} Present address: Park Nicollet Heart Center, Saint Louis Park, Minn.

Journal of Clinical Microbiology, March 2003, p. 1167-1172, Vol. 41, No. 3
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.3.1167-1172.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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