Rapid detection of rifampicin resistance in Mycobacterium tuberculosis by high-resolution melting curve analysis

Malhotra, B. and Goyal, S. and Bhargava, S. and Reddy, P. V. and Chauhan, A. and Tiwari, J. (2015) Rapid detection of rifampicin resistance in Mycobacterium tuberculosis by high-resolution melting curve analysis Int J Tuberc Lung Dis, 19 (12). pp. 1536-1541.

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Abstract

SETTING: High-resolution melting curve analysis (HRMA) can be used to screen for mutations in genes without the need for specific probes, with low turnaround time and high cost-effectiveness. OBJECTIVE: To detect the sensitivity and specificity of a line-probe assay (LPA) and HRMA in comparison with BACTEC™ MGIT™ 960 for the detection of rifampicin (RMP) resistance. DESIGN: A total of 219 Mycobacterium tuberculosis isolates tested by MGIT 960 for RMP susceptibility were tested with HRMA and LPA. Discordant samples were processed for sequencing of the RMP resistance-determining region (RRDR) of the rpoB gene. RESULTS: HRMA identified 93 of 103 (90.3%) isolates that were resistant and 113/116 (97.4%) that were susceptible on MGIT 960, with a sensitivity and specificity of respectively 90.3% and 97.4%. HRMA identified 117/119 (98.3%) LPA-susceptible and 94/100 (94%) resistant isolates, with 98.3% specificity and 94% sensitivity. Two isolates that were susceptible on LPA but resistant on HRMA showed silent mutations at 539 and 541 codons on sequencing, while 6 isolates that were susceptible on HRMA but resistant on LPA showed D516V (n = 4) and H526C/D (n = 2) mutations. Four isolates (3.9%) that were resistant on MGIT were susceptible on all three genotypic methods, which could be due to mutations outside the RRDR or efflux pumps. CONCLUSION: HRMA shows good potential as a rapid screening tool for the detection of drug resistance.

Item Type: Article
Uncontrolled Keywords: Mycobacterium tuberculosis; Genotypic; HRMA; Multidrug-resistant tuberculosis; MDR-TB; RMP resistance
Subjects: Tuberculosis > Laboratory
Tuberculosis > Drug Resistant TB
Institutions: National Task Force, Zonal Task Force & State Task Force of Medical Colleges
Depositing User: Mr SANJAY SINGH
Date Deposited: 21 Dec 2015 09:31
Last Modified: 21 Dec 2015 09:31
URI: http://tbresearch.ntiindia.org.in/id/eprint/2182

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