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... <p>... The third nucleotide (CG<bold>A</bold>) at codon 529, which is a specific nucleotide of <italic>M. tuberculosis</italic> (<xref ref-type="bibr" rid="B8">8</xref>, <xref ref-type="bibr" rid="B11">11</xref>), was not clearly determined by conventional PCR direct sequencing. Furthermore, PCR sequencing misidentified codon 531 as T<bold>C</bold>G, which reflects the rifampin-susceptible strain. However, the second nucleotide at codon 531, which is the most frequent site of mutation related to rifampin resistance, was determined as T<bold>T</bold>G by nested PCR sequencing (Fig. <xref ref-type="fig" rid="F5">5</xref>, lower panel). ...</p> ...