Molecular Characterization of Rifampin-Resistant Isolates of Mycobacterium tuberculosis from Hungary by DNA Sequencing and the Line Probe Assay

doi:10.1128/JCM.39.10.3736-3739.2001

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Journal of Clinical Microbiology, October 2001, p. 3736-3739, Vol. 39, No. 10
0095-1137/01/$04.00+0 DOI: 10.1128/JCM.39.10.3736-3739.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Molecular Characterization of Rifampin-Resistant Isolates of Mycobacterium tuberculosis from Hungary by DNA Sequencing and the Line Probe Assay

Zoltán Bártfai,¹ Ákos Somoskövi,¹^,²^,^* Csaba Ködmön,¹ Nóra Szabó,³^,⁴ Erzsébet Puskás,⁵ Lászlóné Kosztolányi,⁵ Eszter Faragó,⁶ Judit Mester,⁴ Linda M. Parsons,² and Max Salfinger²^,⁷^,⁸

Department of Respiratory Medicine, School of Medicine, Semmelweis University,¹ and Korányi National Institute for Tuberculosis and Respiratory Medicine,⁴ Budapest, Prodia Laboratory for Mycobacteria, Jósa Hospital, Nyíregyháza,³ Borsod-Abaúj-Zeplén County Bureau of Public Health and Medical Officers, Miskolc,⁵ and Laboratory for Mycobacteria, School of Medicine, University of Debrecen, Debrecen,⁶ Hungary, and Wadsworth Center, New York State Department of Health,² Department of Medicine, Albany Medical College,⁷ and Department of Biomedical Sciences, School of Public Health, State University of New York at Albany,⁸ Albany, New York

Received 24 January 2001/Returned for modification 2 May 2001/Accepted 30 May 2001

Two regions of rpoB associated with rifampin resistance were sequenced in 29 rifampin-resistant (determined by the proportionmethod) isolates of Mycobacterium tuberculosis obtained from patientsfrom three counties in Hungary. Of the 29 resistant strains, 27had a mutation in either the 81-bp region (26 strains) or theN-terminal region (1 strain), while the other 2 strains had nomutations in either region. The locations and frequencies of themutations differed from those previously reported. The most commonmutation in this study, D516V, was found in 38% of the Hungarianstrains, a frequency 2 to 10 times higher than that found in studiesfrom other countries. These same 29 isolates were also evaluatedwith the Inno-LiPA Rif. TB test (LiPA), a reverse hybridizationassay for the rapid detection of rifampin resistance. AlthoughLiPA detected the presence of an rpoB mutation in 26 of the resistantisolates, the type of mutation could not be determined in 4 isolatesbecause the mutations present were not among those included onthe LiPA strip. In addition, a silent mutation in one of the rifampin-susceptiblecontrol strains was interpreted as rifampin resistant by LiPA.These findings demonstrate the importance of validating this rapidmolecular test by comparison with DNA sequence results in eachgeographic location before incorporating the test into routinediagnosticwork.

^* Corresponding author. Mailing address: Wadsworth Center, New York State Department of Health, P.O. Box 509, Albany, NY 12201-0509.Phone: (518) 474-2196. Fax: (518) 474-6964. E-mail: somoskov{at}wadsworth.orgor medve{at}pulm.sote.hu.

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