FIELD: biotechnology.
SUBSTANCE: method of detecting the mutations in rpoB gene by carrying out PCR is proposed, when in "real-time" the fragment of rpoB gene is created using primers Rpo11 (5'-GACGTCGAGGCGATCACAC-3') and Rpo12 (5'-GGCACGCTCACTTGACAGAC-3'), after that HRM-analysis is carried out. To increase the reliability of HRM-analysis of mutations in rpoB gene the equal amounts of tested DNA and DNA H37Rv are mixed. The concentration equalisation is carried out using quantitative PCR with use of the same pair of primers with creation of a calibration curve and use of regression equation.
EFFECT: method due to its high specificity and sensitivity enables to identify reliably and quickly the type of mutations in the rpoB gene associated with the emergence of resistance to rifampicin.
2 dwg
