This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Templeton, K. E. Articles by Claas, E. C. J. Search for Related Content PubMed PubMed Citation Articles by Templeton, K. E. Articles by Claas, E. C. J.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, September 2003, p. 4016-4021, Vol. 41, No. 9
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.9.4016-4021.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Development and Clinical Evaluation of an Internally Controlled, Single-Tube Multiplex Real-Time PCR Assay for Detection of Legionella pneumophila and Other Legionella Species

Department of Medical Microbiology, Center of Infectious Diseases, Leiden University Medical Center, Leiden,¹ BioMerieux, Boxtel, The Netherlands,² Department of Microbiology, University of Antwerp, Antwerp, Belgium³

Received 3 March 2003/ Returned for modification 14 April 2003/ Accepted 27 May 2003

A multiplex real-time PCR assay for detection of Legionella pneumophila and Legionella spp. and including an internal control was designed. Legionella species, L. pneumophila, and the internal control were detected simultaneously by probes labeled with 6-carboxy-fluorescein, hexachlorofluorescein, and indodicarbocyanine, respectively. Therefore, no postamplification analysis was required in order to distinguish the targets. The sensitivity of both assays was 2.5 CFU/ml, and from analysis of 10 culture-positive and 74 culture-negative samples from patients investigated for legionellosis, 100% agreement was observed by both assays in comparison to culture. Four additional positives were found by the multiplex real-time PCR assay in the Legionella culture-negative samples.

This article has been cited by other articles:

• Diederen, B. M. W., Kluytmans, J. A. J. W., Vandenbroucke-Grauls, C. M., Peeters, M. F. (2008). Utility of Real-Time PCR for Diagnosis of Legionnaires' Disease in Routine Clinical Practice. J. Clin. Microbiol. 46: 671-677 [Abstract] [Full Text]  
• Bencini, M. A., van den Brule, A. J. C., Claas, E. C. J., Hermans, M. H. A., Melchers, W. J. G., Noordhoek, G. T., Salimans, M. M. M., Schirm, J., Vink, C., van der Zee, A., Jansen, R. (2007). Multicenter Comparison of Molecular Methods for Detection of Legionella spp. in Sputum Samples. J. Clin. Microbiol. 45: 3390-3392 [Abstract] [Full Text]  
• Morozumi, M., Nakayama, E., Iwata, S., Aoki, Y., Hasegawa, K., Kobayashi, R., Chiba, N., Tajima, T., Ubukata, K., the Acute Respiratory Diseases Study Group, (2006). Simultaneous Detection of Pathogens in Clinical Samples from Patients with Community-Acquired Pneumonia by Real-Time PCR with Pathogen-Specific Molecular Beacon Probes. J. Clin. Microbiol. 44: 1440-1446 [Abstract] [Full Text]  
• Espy, M. J., Uhl, J. R., Sloan, L. M., Buckwalter, S. P., Jones, M. F., Vetter, E. A., Yao, J. D. C., Wengenack, N. L., Rosenblatt, J. E., Cockerill, F. R. III, Smith, T. F. (2006). Real-Time PCR in Clinical Microbiology: Applications for Routine Laboratory Testing. Clin. Microbiol. Rev. 19: 165-256 [Abstract] [Full Text]  
• Dirven, K., Ieven, M., Peeters, M. F, van der Zee, A., De Schrijver, K., Goossens, H. (2005). Comparison of three Legionella urinary antigen assays during an outbreak of legionellosis in Belgium. J Med Microbiol 54: 1213-1216 [Abstract] [Full Text]  
• Yanez, M. A., Carrasco-Serrano, C., Barbera, V. M., Catalan, V. (2005). Quantitative Detection of Legionella pneumophila in Water Samples by Immunomagnetic Purification and Real-Time PCR Amplification of the dotA Gene. Appl. Environ. Microbiol. 71: 3433-3441 [Abstract] [Full Text]  
• Ginevra, C., Barranger, C., Ros, A., Mory, O., Stephan, J.-L., Freymuth, F., Joannes, M., Pozzetto, B., Grattard, F. (2005). Development and Evaluation of Chlamylege, a New Commercial Test Allowing Simultaneous Detection and Identification of Legionella, Chlamydophila pneumoniae, and Mycoplasma pneumoniae in Clinical Respiratory Specimens by Multiplex PCR. J. Clin. Microbiol. 43: 3247-3254 [Abstract] [Full Text]  
• Claas, E. C. J., Schilham, M. W., de Brouwer, C. S., Hubacek, P., Echavarria, M., Lankester, A. C., van Tol, M. J. D., Kroes, A. C. M. (2005). Internally Controlled Real-Time PCR Monitoring of Adenovirus DNA Load in Serum or Plasma of Transplant Recipients. J. Clin. Microbiol. 43: 1738-1744 [Abstract] [Full Text]  
• Raggam, R. B., Leitner, E., Berg, J., Muhlbauer, G., Marth, E., Kessler, H. H. (2005). Single-Run, Parallel Detection of DNA from Three Pneumonia-Producing Bacteria by Real-Time Polymerase Chain Reaction. J. Mol. Diagn. 7: 133-138 [Abstract] [Full Text]  
• Roth, S. B., Jalava, J., Ruuskanen, O., Ruohola, A., Nikkari, S. (2004). Use of an Oligonucleotide Array for Laboratory Diagnosis of Bacteria Responsible for Acute Upper Respiratory Infections. J. Clin. Microbiol. 42: 4268-4274 [Abstract] [Full Text]