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Journal of Clinical Microbiology, June 2007, p. 1920-1926, Vol. 45, No. 6
0095-1137/07/$08.00+0     doi:10.1128/JCM.00147-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Assessment of Fluorescent In Situ Hybridization and PCR-Based Methods for Rapid Identification of Burkholderia cepacia Complex Organisms Directly from Sputum Samples ^,

Centre for Infectious Diseases, University of Edinburgh, Edinburgh, United Kingdom

Received 20 January 2007/ Returned for modification 2 March 2007/ Accepted 17 April 2007

Several species within the Burkholderia cepacia complex (BCC) have emerged as significant opportunistic pathogens of patients with cystic fibrosis (CF). BCC infection is typically associated with a poor clinical prognosis and decreased survival. These factors, combined with the existence of highly transmissible epidemic strains, have resulted in strict segregation of BCC- and non-BCC-infected patients to minimize cross infection. Accurate and rapid diagnosis of infections is essential to enable appropriate patient management. However, the rapidly evolving taxonomy of BCC poses a considerable challenge to diagnostics. In the present study, we assessed a commercially available fluorescent in situ hybridization (FISH) assay (seaFAST Cystic Fibrosis I kit) and a novel rRNA gene-based PCR assay for the rapid identification of BCC-positive sputa, irrespective of the BCC species. We report that, while the FISH assay fails to identify all BCC species, it does identify the majority of species, including the two most clinically relevant species, B. multivorans and B. cenocepacia. The sensitivity of the assay applied to sputum was limited by nonspecific background fluorescence. While sputum processing was optimized to minimize background, the resulting sensitivity for BCC detection was 8 x 10⁵ CFU/ml. In contrast, the novel PCR assay reported herein exhibits 100% sensitivity and specificity for all BCC species and can detect 10⁴ CFU/ml when applied to sputum. This novel rRNA gene-based assay is currently the most sensitive BCC-specific PCR assay for the detection of BCC direct from clinical samples and as such is a valuable addition to the field of BCC diagnostics.

Published ahead of print on 25 April 2007.

Supplemental material for this article may be found at http://jcm.asm.org/.

This article has been cited by other articles:

• Lynch, K. H., Dennis, J. J. (2008). Development of a Species-Specific fur Gene-Based Method for Identification of the Burkholderia cepacia Complex. J. Clin. Microbiol. 46: 447-455 [Abstract] [Full Text]