Single strand conformation polymorphism profiles with biotinylated PCR products to detect mutations in rpoB gene of Mycobacterium tuberculosis

Selvakumar, N and Wilson, S M and McNerney, R and Narayanan, P R (1997) Single strand conformation polymorphism profiles with biotinylated PCR products to detect mutations in rpoB gene of Mycobacterium tuberculosis. Current Science, 73 (10). pp. 774-777. ISSN 0011-3891

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Abstract

A fragment of the rpoB gene, including the region shown to be involved in rifampicin resistance, was amplified from 15 rifampicin-resistant and 6 rifampicin- sensitive clinical isolates of Mycobacterium tuberculosis by the polymerase chain reaction (PCR). One of the primers, employed in PCR, was biotinylated. The biotinylated strand of the PCR product was separated from the unbiotinylated strand using streptavidin magnetic beads. Both the strands were subjected to single strand conformation polymorphism in polyacrylamide gel electrophoresis. The DNA bands were silver stained to study their migration pattern. A shift in the migration of either of the strands of the test strain compared to the strands from a control rifampicin-sensitive strain was considered as indicative of resistance. This strategy was found to ease the visualization of shift in the migration of the strands in 17 of 21 samples and thereby detection of mutations.

Item Type:	Article
Subjects:	Tuberculosis > Laboratory Research > Bacteriological
Divisions:	Basic Science Research > Bacteriology
Depositing User:	Dr. Rathinasabapati R
Date Deposited:	29 Oct 2013 11:20
Last Modified:	14 Mar 2016 04:50
URI:	http://eprints.nirt.res.in/id/eprint/459

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