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Journal of Clinical Microbiology, July 2003, p. 3384-3386, Vol. 41, No. 7
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.7.3384-3386.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Low-Stringency Single-Specific-Primer PCR as a Tool for Detection of Mutations in the rpoB Gene of Rifampin-Resistant Mycobacterium tuberculosis

Wania S. Carvalho,1* Silvana Spindola de Miranda,2 Kátia M. Costa,3 José G.V.C. Araújo,3 Claudio J. Augusto,4 João B. Pesquero,5 Jorge L. Pesquero,3 and Maria A. Gomes3

Departamento de Farmácia Social, Faculdade de Farmácia,1 Faculdade de Medicina,2 and Instituto de Ciencias Biológicas,3 Fundação Ezequiel Dias, Belo Horizonte, Minas Gerais,4 Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo, Brazil5

Received 4 September 2002/ Returned for modification 6 November 2002/ Accepted 23 March 2003

By the low-stringency single-specific-primer PCR technique, a highly sensitive and rapid method for diagnosis of rifampin resistance in Mycobacterium tuberculosis was established. Seven rifampin-resistant and five rifampin-susceptible specimens were analyzed. Rifampin resistance was determined by MIC measurement. A complex electrophoretic pattern consisting of many bands was obtained for both susceptible and rifampin-resistant isolates. The same pattern was obtained for all of the susceptible specimens, but differences between resistant and susceptible isolates were found. DNA sequencing showed that a particular mutation produces a specific electrophoretic pattern.

* Corresponding author. Mailing address: Departamento de Farmácia Social, Faculdade de Farmácia, Universidade Federal de Minas Gerais, Av. Olegário Maciel, 2360, 30180-112, Belo Horizonte, MG, Brazil. Phone: 55-31-3499-2942. Fax: 55-31-3339-7685. E-mail: wscarv{at}icb.ufmg.br.

Journal of Clinical Microbiology, July 2003, p. 3384-3386, Vol. 41, No. 7
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.7.3384-3386.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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