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7% [20] and 67.6% [21] of all INH-resistant strains identified. This represents >30% of INH-resistant strains with the codon 315 mutation detected in the katG gene. Therefore, our results emphasize the importance of studying genes related to INH resistance other than katG. Previous reports have described the application of pyrosequencing to the identification of RMP-resistant M.?tuberculosis isolates [22�C26]. The advantage of the method described here compared with previous studies is the possibility of including molecular monitoring LY294002 solubility dmso as well as analysis of specific mutations. This is supported by the fact that in the evaluation phase, this method was able to detect mutations that were not present in the development of the assay. These mutations were both in the rpoB (CAGCAA mutation in codon 517, and GAC GTC transition in codon 516) and inhA (?15 CT) genes. In the analysis of all strains [n?=?171: development (n?=?75) plus evaluations (n?=?96)], the most frequent site for mutations in M.?tuberculosis strains was in codon 531 of the rpoB gene (in 18 out of 41 resistant strains). The two base pair substitutions at this codon were TCGTTG and TCGTGG. The second most common site for mutations was in codon 526, where the wild-type CAC codon was replaced by either TGC, GAC or CGC (in nine out of 41 resistant strains). The high frequency of mutation at codon 531 in RMP-resistant this website strains, especially the TGCTTG substitution, is consistent with other reports that have analysed the same region in a variety of populations [11,15,16]. In contrast, no mutations were detected in 12 of the 41 RMP-resistant strains analysed. We hypothesize that for this subset of isolates, the mutations associated with RMP resistance are present either in the RRDR region not monitored in this study (codon 507�C514), outside the RRDR, or in other genes. The role of S6 Kinase unknown mechanisms in drug resistance, independent of or in conjunction with genetic mutations, cannot be excluded. Additional data extracted from the analysis of all 171 strains showed a correlation linking mutations in ahpC and inhA with resistance to INH (p?
