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Journal of Clinical Microbiology, February 2007, p. 347-350, Vol. 45, No. 2
0095-1137/07/$08.00+0     doi:10.1128/JCM.01303-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Detection and Differentiation of Bordetella spp. by Real-Time PCR{triangledown}

Christoph Koidl,1 Michael Bozic,1 Anja Burmeister,2 Markus Hess,2 Egon Marth,1 and Harald H. Kessler1*

Institute of Hygiene, Medical University of Graz, Graz, Austria,1 QIAGEN Hamburg GmbH, Research and Development, Hamburg, Germany2

Received 26 June 2006/ Returned for modification 14 September 2006/ Accepted 22 November 2006

Molecular detection of Bordetella pertussis DNA is a sensitive and specific method for the rapid diagnosis of pertussis. In this study, a new molecular assay for the detection and differentiation of Bordetella spp. based on automated DNA extraction and real-time PCR was evaluated. The analytical sensitivity of the new assay was determined by Probit analysis of serial dilutions of both cloned PCR products IS481 and IS1001 and cell suspensions of B. pertussis, B. parapertussis, and B. bronchiseptica. The specificity was analyzed by testing a number of pathogens producing respiratory infections. Moreover, a total of 92 clinical samples were investigated. The results were compared to those obtained by an in-house assay based on manual DNA extraction, followed by real-time PCR and detection of IS481. The analytical sensitivity of the new assay for the detection of IS481 and IS1001 was determined to be 2.2 and 1.2 genome equivalents/µl, respectively. The analytical sensitivity for the detection of B. pertussis, B. parapertussis, and B. bronchiseptica was determined to be 1.6, 1.0, and 2.7 genome equivalents/µl, respectively. When clinical specimens were tested with the new assay, 46 of 92 were found to be positive for Bordetella DNA. With the in-house assay, 45 samples tested positive. The new molecular assay proved to be suitable for the rapid diagnosis of pertussis in the routine diagnostic laboratory.

* Corresponding author. Mailing address: Institute of Hygiene, Medical University of Graz, Universitaetsplatz 4, 8010 Graz, Austria. Phone: 43 (316) 380-4383. Fax: 43 (316) 380-9648. E-mail: harald.kessler{at}meduni-graz.at.

{triangledown} Published ahead of print on 6 December 2006.

Journal of Clinical Microbiology, February 2007, p. 347-350, Vol. 45, No. 2
0095-1137/07/$08.00+0     doi:10.1128/JCM.01303-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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