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Free Content Performance comparison of four methods for detecting multidrug-resistant Mycobacterium tuberculosis strains

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SETTING: National Tuberculosis Reference Laboratory, Kuwait.

OBJECTIVE: To compare Genotype MTBDRplus (gMTBDR+), INNO-LiPA Rif.TB (INNO-LiPA), polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and DNA sequencing for detecting rifampicin (RMP) and/or isoniazid (INH) resistance-associated mutations in the rpoB hot-spot region (HSR-rpoB), the katG codon 315 (katG315) and the inhA regulatory region (inhA-RR) among multidrug-resistant Mycobacterium tuberculosis (MDR-TB) isolates.

DESIGN: A total of 82 MDR-TB and 43 pansusceptible M.tuberculosis BACTEC 460-characterised isolates were processed using molecular techniques and the Mycobacterial Growth Indicator Tube (MGIT) 960 system.

RESULTS: All susceptible strains contained wild-type sequences in target genes. RMP resistance was detected in respectively 78, 77 and 79 MDR-TB strains by gMTBDR+, INNO-LiPA and HSR-rpoB sequencing. Two isolates with Ins514TTC mutation were detected as RMP-resistant by gMTBDR+ but as RMP-susceptible by INNO-LiPA. One isolate with L533P mutation, detected as RMP-susceptible by gMTBDR+, was detected as RMP-resistant by INNO-LiPA. Two of three isolates detected as RMP-susceptible by gMTBDR+, INNO-LiPA, HSR-rpoB sequencing and the MGIT 960 system contained a I572F mutation that is outside HSR-rpoB. INH resistance was detected in respectively 76, 60, 60 and 22 MDR-TB strains by gMTBDR+, katG315 PCR-RFLP, katG315 sequencing and inhA-RR sequencing.

CONCLUSIONS: Although gMTBDR+ accurately detected ∼88% of MDR-TB strains, some rpoB mutations were either missed or were outside the region of analysis of the gMTBDR+ assay.

Keywords: Mycobacterium tuberculosis; isoniazid; molecular detection; multidrug resistance; rifampicin

Document Type: Regular Paper

Affiliations: Department of Microbiology, Faculty of Medicine, Kuwait University, Kuwait

Publication date: January 1, 2011

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  • The International Journal of Tuberculosis and Lung Disease publishes articles on all aspects of lung health, including public health-related issues such as training programmes, cost-benefit analysis, legislation, epidemiology, intervention studies and health systems research. The IJTLD is dedicated to the continuing education of physicians and health personnel and the dissemination of information on tuberculosis and lung health world-wide.

    Certain IJTLD articles are selected for translation into French, Spanish, Chinese or Russian. They are available on the Union website

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