The aim was to evaluate the Single-Strand Conformation Polymorphism method as a potential tool for rapid detection of rifampicin-resistant strains by the use of 39 rifampicin-resistant Mycobacterium tuberculosis strains isolated in Serbia. SSCP analysis on acrylamide gel detected 56.4% of the rifampicin-resistant M. tuberculosis strains and showed the inability to detect one of the most frequent mutations, TCG-->TTG mutation in codon 531 of the rpoB gene, which was shown by automated sequencing.