Performance comparison of four methods for detecting multidrug-resistant Mycobacterium tuberculosis strains
Abstract: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.
Document Type: Regular Paper
Affiliations: Department of Microbiology, Faculty of Medicine, Kuwait University, Kuwait
Publication date: January 1, 2011
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