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Journal of Clinical Microbiology, January 2001, p. 107-110, Vol. 39, No. 1
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.1.107-110.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Frequency of rpoB Mutations Inside and Outside the Cluster I Region in Rifampin-Resistant Clinical Mycobacterium tuberculosis Isolates

Markus Heep,1,* Barbara Brandstätter,1 Ulrich Rieger,1 Norbert Lehn,1 Elvira Richter,2 Sabine Rüsch-Gerdes,2 and Stefan Niemann2

Institut fuer Medizinische Mikrobiologie und Hygiene, Universität Regensburg, Regensburg,1 and Forschungszentrum Borstel, National Reference Center for Mycobacteria, Borstel,2 Germany

Received 28 August 2000/Returned for modification 25 September 2000/Accepted 31 October 2000

The prevalence of recently described mutation V176F, located in the beginning of the rpoB gene and associated with rifampin resistance and the wild-type cluster I sequence, was determined by analyzing the distribution of rpoB mutations among 80 rifampin (RIF)-resistant Mycobacterium tuberculosis strains isolated in Germany during 1997. The most frequent rpoB mutations were changes in codon 456 (52 isolates, 65%), followed by changes in codon 441 (13 isolates, 16%) and codon 451 (11 isolates, 14%). The V176F mutation was detected in one isolate of the study population and in 5 of 18 RIF-resistant strains with no cluster I mutation from six previously published studies. In three isolates, a mixture of resistant and susceptible subpopulations (heteroresistance) prohibited the detection of rpoB mutations in the initial analysis; however, in these isolates, cluster I mutations could be verified after a passage on RIF-containing medium. IS6110 DNA fingerprinting of 76 strains revealed eight clusters comprising 27 strains with identical restriction fragment length polymorphism patterns that mainly also show identical rpoB mutations and identical or similar drug resistance patterns. In conclusion, our results indicate that the V176F mutation should be included in molecular tests for prediction of RIF resistance in M. tuberculosis. We further demonstrated that heteroresistance caused by a mixture of mycobacterial subpopulations with different susceptibilities to RIF may influence the sensitivity of molecular tests for detection of resistance.

* Corresponding author. Mailing address: Institut für Medizinische Mikrobiologie und Hygiene, Universität Freiburg, Herrman-Herder-Str-11, D-79104 Freiburg, Germany. Phone: 49 761 2036546. Fax: 49 761 2036562. E-mail: Markus-Heep{at}web.de.

Journal of Clinical Microbiology, January 2001, p. 107-110, Vol. 39, No. 1
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.1.107-110.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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