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Journal of Clinical Microbiology, July 1998, p. 1977-1983, Vol. 36, No. 7
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Genetic Diversity among Strains of Moraxella catarrhalis: Analysis Using Multiple DNA Probes and a Single-Locus PCR-Restriction Fragment Length Polymorphism Method

Elaine S. Walker,1,2,* Robert A. Preston,3 J. Christopher Post,3 Garth D. Ehrlich,3 John H. Kalbfleisch,4 and Karin L. Klingman5

James H. Quillen Veterans Affairs Medical Center,1 Department of Internal Medicine,2 and Department of Medical Education,4 Quillen College of Medicine, East Tennessee State University, Johnson City, Tennessee; Center for Genomic Sciences, Department of Otolaryngology, Allegheny University of Health Sciences, Pittsburgh, Pennsylvania3; and Buffalo Veterans Affairs Medical Center and Department of Medicine, State University of New York, Buffalo, New York5

Received 4 November 1996/Returned for modification 31 January 1997/Accepted 10 April 1998

Moraxella (Branhamella) catarrhalis, a causative agent of otitis media, sinusitis, and exacerbation of bronchitis, has acquired widespread ability to produce beta -lactamase and can be nosocomially transmitted. The typing methods used in epidemiological analyses of M. catarrhalis are not optimal for genetic analyses. Two methods, a multiple-locus Southern blot (SB) method and a single-locus PCR-restriction fragment length polymorphism (RFLP) method, were developed and used to assess genetic diversity and potential clinical and geographic relationships in M. catarrhalis. Nine randomly cloned M. catarrhalis DNA fragments were used as probes of SBs containing DNA from 54 geographically and clinically diverse strains. For comparison, a PCR-RFLP method was developed as a quick, inexpensive, and discriminating alternative. A highly variable 3.7-kb genomic region (M46) was cloned and sequenced, and 3.5 kb of the cloned DNA was targeted for PCR amplification. DNAs from the 54 strains were subjected to PCR-RFLP. SB analysis distinguished all strains that had no apparent epidemiological linkage (40 of 54), and PCR-RFLP distinguished fewer strains (21 of 54). Epidemiologically linked strains appeared genetically identical by both methods. PCR-RFLP was compared to pulsed-field gel electrophoresis (PFGE) for 8 of the 54 strains and 23 additional strains. PCR-RFLP distinguished fewer strains than PFGE typing (16 of 31 versus 20 of 31 strains), but PCR-RFLP was more useful for inferring interstrain relatedness. Separate cluster analyses of multilocus SB and single locus PCR-RFLP data showed high genetic diversity within and across geographic locations and clinical presentations. The resultant dendrograms were not entirely concordant, but both methods often gave similar strain clusters at the terminal branches. High genetic diversity, nonconcordance of cluster analyses from different genetic loci, and shared genotypes among epidemiologically linked strains support a hypothesis of high recombination relative to spread of clones. Single-locus PCR-RFLP may be suitable for short-term epidemiological studies, but the SB data demonstrate that greater strain discrimination may be obtained by sampling variation at multiple genomic sites.

* Corresponding author. Mailing address: Department of Veterans Affairs Medical Center (11C), Johnson City, P.O. Box 4000, Mountain Home, TN 37684-4000. Phone: (423) 439-8333. Fax: (423) 232-6392. E-mail: walker.elaine{at}mtn-home.va.gov.

Journal of Clinical Microbiology, July 1998, p. 1977-1983, Vol. 36, No. 7
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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