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Journal of Clinical Microbiology, July 2005, p. 3247-3254, Vol. 43, No. 7
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.7.3247-3254.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

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

C. Ginevra,1,2 C. Barranger,2 A. Ros,1 O. Mory,3 J.-L. Stephan,3 F. Freymuth,4 M. Joannès,2 B. Pozzetto,1 and F. Grattard1*

Laboratoire de Bactériologie-Virologie, GIMAP, Faculté de Médecine Jacques Lisfranc, Saint-Etienne, France,1 Argene Inc., North Massapequa, New York 11758,2 Service de Pédiatrie, Centre Hospitalier Universitaire, Saint-Etienne, France,3 Laboratoire de Virologie, Centre Hospitalier Universitaire, Caen, France4

Received 15 November 2004/ Returned for modification 24 January 2005/ Accepted 28 February 2005

This study describes the development and evaluation of a new commercial test, Chlamylege (Argene Inc.), which allows the simultaneous detection in respiratory samples of Chlamydophila pneumoniae, Mycoplasma pneumoniae, and most Legionella species, as well as PCR inhibitors, by using a multiplex PCR and microplate hybridization. The sensitivities of Chlamylege were 1 x 10–3 IFU, 5 x 10–2 color-changing units, and 1 CFU per reaction tube for C. pneumoniae, M. pneumoniae, and Legionella pneumophila, respectively. A cohort of 154 clinical samples from patients with documented respiratory infections was analyzed by the kit, including 2 samples from patients with C. pneumoniae infection, 9 samples from patients with M. pneumoniae infection, 19 samples from patients with Legionella species infection, and 114 samples that tested negative for the three pathogens. All the positive specimens were correctly detected and identified by the Chlamylege kit, and no false-positive result was observed with the negative samples. The kit was then evaluated in a pediatric prospective study that included 220 endotracheal aspirates, and the results were compared with those obtained by three single in-house PCR assays. Four specimens were found to be positive for C. pneumoniae and six were found to be positive for M. pneumoniae by using both strategies. The Chlamylege kit detected two additional samples positive for M. pneumoniae and one additional sample positive for a Legionella species other than L. pneumophila; these three samples were shown to be true positive by other techniques. These overall results demonstrate that the Chlamylege assay is sensitive, specific, and convenient for the rapid detection and identification of atypical pathogens in clinical samples from patients with respiratory infections.

* Corresponding author. Mailing address: Laboratoire de Bactériologie-Virologie, Hôpital Nord, CHU de Saint-Etienne, 42055 Saint-Etienne cedex 02, France. Phone: 33 4 77 82 83 15. Fax: 33 4 77 82 84 60. E-mail: florence.grattard{at}univ-st-etienne.fr.

Journal of Clinical Microbiology, July 2005, p. 3247-3254, Vol. 43, No. 7
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.7.3247-3254.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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