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Indian Journal of Experimental Biology (IJEB) >
IJEB Vol.44 [2006] >
IJEB Vol.44(07) [July 2006] >
| Title: | Molecular characterization of mutation associated with rifampicin and isoniazid resistance in Mycobacterium tuberculosis isolates |
| Authors: | Negi, Sanjay Singh
Anand, Ranjana
Pasha, S T
Gupta, Sunil
Basir, Seemi Farhat
Khare, Shashi
Lal, Shiv |
| Keywords: | Isoniazid
Mycobacterium tuberculosis
Rifampicin
Tuberculosis |
| Issue Date: | Jul-2006 |
| Publisher: | CSIR |
| Abstract: | Nucleotide changes in catalase peroxidase (Kat G) gene and gene encoding the beta
subunit of RNA polymerase (rpo B),
responsible for isoniazid and rifampicin drug resistance were determined in the
clinical isolates of Mycobacterium
tuberculosis by PCR-RFLP, Line probe assay and DNA sequencing. PCR-RFLP test was performed by HapII
cleavage of an amplified fragment of Kat
G gene to detect the transversion 315AGC→ACC(Ser→Thr)
which is associated with INH drug resistance. The Line probe assay kit was
evaluated to detect the mutation in 81bp RMP resistance determining region of rpo B gene associated with RMP drug
resistance. These results were validated by DNA sequencing and drug
susceptibility test. Kat G S 315 T mutation was found in
74.19% strains of M. tuberculosis
from Delhi. This mutation was not found in any of the susceptible strains
tested. The line probe assay kit and
DNA sequencing identified 18 isolates
as RMP resistant with specific mutation, while
one of the RMP resistant strain was identified as RMP susceptible, with a
concordance of 94.73% with the phenotypic
drug susceptibility result. Majority
(8 of 19, 42.1%) of resistant isolates involved base changes at codon 531 of rpo
B gene. Both PCR-RFLP and Line
probe assay test can be used in many of the clinical microbiology laboratories
for early detection of isoniazid and rifampicin drug resistance in clinical
isolates of M. tuberculosis. |
| Page(s): | 547-553 |
| ISSN: | 0975-1009 (Online); 0019-5189 (Print) |
| Source: | IJEB Vol.44(07) [July 2006]
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