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Journal of Clinical Microbiology, January 2003, p. 295-303, Vol. 41, No. 1
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.1.295-303.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Culture-Independent Molecular Analysis of Microbial Constituents of the Healthy Human Outer Ear

Daniel N. Frank,1 George B. Spiegelman,2 William Davis,1 Eileen Wagner,1 Eric Lyons,3 and Norman R. Pace1*

Department of Molecular, Cellular, and Developmental Biology, University of Colorado, Boulder, Colorado 80309-0347,1 Department of Microbiology and Immunology, University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z3,2 Molecular Sciences Institute, Berkeley, California 947043

Received 26 April 2002/ Returned for modification 16 July 2002/ Accepted 24 October 2002

Molecular-phylogenetic sequence analyses have provided a new perspective on microbial communities by allowing the detection and identification of constituent microorganisms in the absence of cultivation. In this study we used broad-specificity amplification of ribosomal DNA (rDNA) genes to survey organisms present in the human outer ear canal. Samples were obtained from 24 individuals, including members of three extended families, in order to survey the resident microbiota and to examine microbial population structures in individuals related by familial or household associations. To examine the stability of the microbial populations, one individual was sampled four times and another twice over a 14-month period. We found that a distinct set of microbial types was present in the majority of the subjects sampled. The two most prevalent rDNA sequence types that were identified in multiple individuals corresponded closely to those of Alloiococcus otitis and Corynebacterium otitidis, commonly thought to be associated exclusively with infections of the middle ear. Our results suggest, therefore, that the outer ear canal may serve as a reservoir for normally commensal microbes that can contribute to pathogenesis upon introduction into the middle ear. Alternatively, culture analyses of diseases of the middle ear may have been confounded by these contaminating commensal organisms.

* Corresponding author. Mailing address: Department of Molecular, Cellular, and Developmental Biology, University of Colorado, Boulder, CO 80309-0347. Phone: (303) 735-1864. Fax: (303) 492-7744. E-mail: nrpace{at}colorado.edu.

Journal of Clinical Microbiology, January 2003, p. 295-303, Vol. 41, No. 1
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.1.295-303.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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