Rapid Detection of Mutation in RRDR of <Emphasis Type="Italic">rpo B</Emphasis> Gene for Rifampicin Resistance in MDR-Pulmonary Tuberculosis by DNA Sequencing |
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| Authors: | Surajeet Kumar Patra Anju Jain B L Sherwal Ashwani Khanna |
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| Institution: | (1) Department of Biochemistry, Lady Hardinge Medical College, New Delhi, India;(2) Department of Microbiology, Lady Hardinge Medical College, New Delhi, India;(3) Department of Chest Clinic, Lok Nayak Hospital, New Delhi, India |
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| Abstract: | To detect the site of mutation in RRDR of rpo B gene for rifampicin resistance in MDR-TB by DNA sequencing. 50 MDR-TB patients
were enrolled in our study after informed written consent. Mycobacterial DNA was extracted from sputum samples by Universal
Sample Processing (USP) method and RRDR of rpo B gene was amplified by PCR using primers RP4T and RP8T and then sequenced
by automated DNA sequencing. The nucleotide sequences of RRDR of rpo B gene were compared with the reference sequence. We
observed three different types of mutation in the RRDR of rpo B gene. The frequency of mutation in codon 531 (TCG → TTG),
526 (CAC → TAC) and 516 (GAC → GTC) are 60, 26.6 and 6.6% respectively. Of the total cases studied, 6.6% cases, although resistant
to rifampicin, did not show any mutation in the RRDR of rpo B gene. Codon 531 (TCG → TTG) is the most common site of mutation
in RRDR of rpo B gene for rifampicin resistance in MDR-pulmonary tuberculosis followed by codon 526 (CAC → TAC) and codon
516 (GAC → GTC). |
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| Keywords: | Multidrug resistance Tuberculosis Rifampicin resistance rpo B gene |
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