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Journal of Clinical Microbiology, May 2007, p. 1588-1593, Vol. 45, No. 5
0095-1137/07/$08.00+0 doi:10.1128/JCM.01963-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Department of Oral Medicine, Carolinas Medical Center, Charlotte, North Carolina,1 Department of Molecular Genetics, The Forsyth Institute, Boston, Massachusetts,2 Division of Respiratory, Critical Care and Occupational Pulmonary Medicine, University of Utah Medical Center, Salt Lake City, Utah3
Received 21 September 2006/ Returned for modification 16 January 2007/ Accepted 5 February 2007
Trauma intensive care unit (TICU) patients requiring mechanical respiratory support frequently develop ventilator-associated pneumonia (VAP). Oral and oropharyngeal bacteria are believed to be responsible for many cases of VAP, but definitive evidence of this relationship is lacking. Earlier studies used conventional culture-based methods for identification of bacterial pathogens, but these methods are insufficient, as some bacteria may be uncultivable or difficult to grow. The purpose of this study was to use a culture-independent molecular approach to analyze and compare the bacterial species colonizing the oral cavity and the lungs of TICU patients who developed VAP. Bacterial samples were acquired from the dorsal tongue and bronchoalveolar lavage fluid of 16 patients. Bacterial DNA was extracted, and the 16S rRNA genes were PCR amplified, cloned into Escherichia coli, and sequenced. The sequencing data revealed the following: (i) a wide diversity of bacterial species in both the oral and pulmonary sites, some of them novel; (ii) known and putative respiratory pathogens colonizing both the oral cavity and lungs of 14 patients; and (iii) a number of bacterial pathogens (e.g., Dialister pneumosintes, Haemophilus segnis, Gemella morbillorum, and Pseudomonas fluorescens) in lung samples that had not been reported previously at this site when culture-based methods were used. Our data indicate that the dorsal surface of the tongue serves as a potential reservoir for bacterial species involved in VAP. Furthermore, it is clear that the diversity of bacterial pathogens for VAP is far more complex than the current literature suggests.
Published ahead of print on 14 February 2007.
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