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Journal of Clinical Microbiology, April 2005, p. 1564-1569, Vol. 43, No. 4
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.4.1564-1569.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Comparison of an Internally Controlled, Large-Volume LightCycler Assay for Detection of Mycobacterium tuberculosis in Clinical Samples with the COBAS AMPLICOR Assay

Siegfried Burggraf,1* Udo Reischl,2 Naeem Malik,1 Markus Bollwein,2 Ludmila Naumann,3 and Bernhard Olgemöller1

Labor Becker, Olgemöller und Kollegen, München,1 Institut für Medizinische Mikrobiologie und Hygiene, University of Regensburg,2 Bayerisches Landesamt für Gesundheit und Lebensmittelsicherheit, Oberschleißheim, Germany3

Received 22 October 2004/ Returned for modification 23 November 2004/ Accepted 17 December 2004

We present a sensitive and specific assay for reliable and flexible detection of members of the Mycobacterium tuberculosis complex (MTBC) in clinical samples. This real-time PCR assay, which uses the LightCycler 2.0 instrument and 100-µl glass capillaries, can provide a result within 1 h after DNA extraction. The primers amplify a 206-bp fragment of the MTBC 16S rRNA gene. The sensor hybridization probe targets a region highly specific to members of the MTBC. The assay also includes a novel type of internal control that monitors the function of the reaction components and can detect potential inhibitors. Template DNA was extracted by the same procedure used for the COBAS AMPLICOR M. tuberculosis assay, so the LightCycler assay could be directly compared to the COBAS AMPLICOR assay. The LightCycler assay was evaluated with 146 clinical samples of various types. Very good agreement (100% sensitivity, 98.6% specificity) could be shown between the LightCycler and COBAS AMPLICOR assays. Specificity was checked with a panel of nontuberculous mycobacteria, as well as a large panel of bacterial and fungal organisms.

* Corresponding author. Mailing address: Labor Becker, Olgemöller und Kollegen, Führichstrasse 70, D-81671 München, Germany. Phone: 49-89-450917-463. Fax: 49-89-450917-300. E-mail: sburggraf{at}labor-bo.de.

Journal of Clinical Microbiology, April 2005, p. 1564-1569, Vol. 43, No. 4
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.4.1564-1569.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Bastien, P., Procop, G. W., Reischl, U. (2008). Quantitative Real-Time PCR Is Not More Sensitive than "Conventional" PCR. J. Clin. Microbiol. 46: 1897-1900 [Full Text]  


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