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Journal of Clinical Microbiology, January 1998, p. 20-23, Vol. 36, No. 1
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Characterization of rpoB Mutations in Rifampin-Resistant Clinical Mycobacterium tuberculosis Isolates from Greece

Peggy Matsiota-Bernard,1,* Georgia Vrioni,1,2 and Evangelos Marinis2

Laboratoire de Microbiologie, Hôpital Raymond Poincaré, 92380 Garches, France,1 and Reference Center for Tuberculosis, Laboratory of Microbiology, Sotiria Hospital, Athens, Greece2

Received 26 June 1997/Returned for modification 1 August 1997/Accepted 6 October 1997

There is a geographic distribution of Mycobacterium tuberculosis strains with various rpoB gene mutations that account for rifampin resistance. We studied 17 rifampin-resistant clinical isolates from patients in Greece to identify rpoB mutations. The aim of our study was the evaluation of a commercially available line probe assay kit (INNO-LiPA Rif. TB) to detect rpoB mutations and rifampin resistance. The results obtained with the commercially available assay were compared to those obtained by automated DNA sequence analysis of amplified PCR products. Randomly amplified polymorphic DNA (RAPD) analyses of the isolates were also performed. The overall concordance of the line probe assay with phenotypic rifampin susceptibility test was 94%. Three distinct rpoB mutations in codons Ser531, His526, and Asp516 were correctly identified with the kit, but mutations in external regions and insertions were detected only by automated DNA sequence analysis. The changes in codons Ser531 and His526 accounted for the majority of rifampin resistance, as previously described for isolates from other geographic areas. The results obtained by RAPD analyses of the isolates suggested that clonally related M. tuberculosis strains can have subclones bearing distinct mutant rpoB alleles. We conclude that this line probe assay kit, which is fast and with which tests are easy to perform, can be used for the rapid detection of rifampin resistance in M. tuberculosis before the availability of results by conventional methods and for epidemiological studies but that negative results obtained by this method do not rule out rifampin resistance.

* Corresponding author. Mailing address: Laboratoire de Microbiologie, Hôpital Raymond Poincaré, 104, bd Raymond Poincaré, 92380 Garches, France. Phone: (1) 47 10 79 50. Fax: (1) 47 10 79 49. E-mail: peggy.matsiota-bernard{at}rpc.ap-hop-paris.fr.

Journal of Clinical Microbiology, January 1998, p. 20-23, Vol. 36, No. 1
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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