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Journal of Clinical Microbiology, November 2009, p. 3461-3465, Vol. 47, No. 11
0095-1137/09/$08.00+0     doi:10.1128/JCM.01730-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Multicenter Evaluation of a Transcription-Reverse Transcription Concerted Assay for Rapid Detection of Mycobacterium tuberculosis Complex in Clinical Specimens {triangledown}

V. Drouillon,1 G. Delogu,2 G. Dettori,3 P. H. Lagrange,1 M. Benecchi,3 F. Houriez,1 K. Baroli,1 F. Ardito,2 R. Torelli,2 C. Chezzi,3 G. Fadda,2 and J.-L. Herrmann1,4*

Assistance Publique-Hôpitaux de Paris, Microbiology Laboratory, Saint Louis Hospital, 1 Avenue Claude Vellefaux, 75475 Paris Cedex 10,1 Assistance Publique-Hôpitaux de Paris, Microbiology Laboratory, Raymond Poincaré Hospital, 104 Boulevard Raymond Poincaré, 92380 Garches, France,4 Universita Cattolica Del Sacro Cuore, Institute of Microbiology, Largo F. Vito, 00168 Rome,2 University Hospital Parma, Section of Microbiology, Via Gramsci 14, 43100 Parma, Italy3

Received 8 September 2008/ Returned for modification 26 May 2009/ Accepted 30 August 2009

A European multicenter study was performed to evaluate the performance of a new method, based on the transcription-reverse transcription concerted reaction (TRC-2), which enabled one-step amplification and real-time detection of the Mycobacterium tuberculosis 16S rRNA target directly in clinical specimens. A total of 633 respiratory and nonrespiratory specimens were tested, and the results were compared with those from smears and cultures. A total of 129 patients (Paris center) were followed up in order to evaluate the clinical performance of TRC-2. By using M. tuberculosis complex strains to inoculate sterile sputa, the detection limit of TRC-2 was found to be 30 to 50 CFU/ml. A total of 548 respiratory specimens and 59 extrapulmonary specimens were assessable. For pulmonary specimens, the sensitivities of TRC-2 and acid-fast smear were 86.8% and 50.4%, respectively (P = 0.002). The specificities were 97.5% and 100%, respectively. For extrapulmonary specimens, the sensitivities of TRC-2 and acid-fast smear were 83.3% and 8.3% (P < 0.0001), and the specificities were 95.8% and 100%, respectively. Fifteen of 129 patients were diagnosed with pulmonary tuberculosis (TB). The sensitivities of culture and TRC-2 were 80% (12/15) and 86.7% (13/15) (P = 0.16), and the specificities were 100% and 93.9%, respectively. Based on an 11.6% incidence of TB in our population, the positive predictive values of TRC-2 and culture were 81.3% and 100%, respectively, and the negative predictive values were 98.2% and 97.4%, respectively. These results demonstrated that detection of M. tuberculosis complex in clinical specimens by TRC-2 with ready-to-use reagents was an efficient and rapid method for the diagnosis of pulmonary and extrapulmonary TB.

* Corresponding author. Mailing address: Assistance Publique-Hôpitaux de Paris, Raymond Poincaré Hospital, Microbiology Laboratory, 104 Boulevard Raymond Poincaré, 92380 Garches, France. Phone: 33147107950. Fax: 33147107949. E-mail: jean-louis.herrmann{at}rpc.aphp.fr

{triangledown} Published ahead of print on 9 September 2009.

Journal of Clinical Microbiology, November 2009, p. 3461-3465, Vol. 47, No. 11
0095-1137/09/$08.00+0     doi:10.1128/JCM.01730-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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