Author:

Keywords:

Science & Technology, Life Sciences & Biomedicine, Microbiology, MYCOBACTERIUM-TUBERCULOSIS, ETHAMBUTOL RESISTANCE, FUNCTIONAL-ANALYSIS, GYRASE MUTATIONS, MTBDRPLUS ASSAY, STRAINS, PERFORMANCE, OUTCOMES, Antitubercular Agents, DNA Gyrase, DNA, Bacterial, Democratic Republic of the Congo, Disease Outbreaks, False Positive Reactions, Fluoroquinolones, Genotyping Techniques, Humans, Mutation, Missense, Mycobacterium tuberculosis, Point Mutation, Sequence Analysis, DNA, Sputum, Tuberculosis, Multidrug-Resistant, 06 Biological Sciences, 07 Agricultural and Veterinary Sciences, 11 Medical and Health Sciences, 3202 Clinical sciences, 3207 Medical microbiology

Abstract:

Fluoroquinolones are the core drugs for the management of multidrug-resistant tuberculosis (MDR-TB). Molecular drug susceptibility testing methods provide considerable advantages for scaling up programmatic management and surveillance of drug-resistant TB. We describe here the misidentification of fluoroquinolone resistance by the GenoType MTBDRsl (MTBDRsl) (Hain Lifescience GmbH, Nehren, Germany) line probe assay (LPA) encountered during a feasibility and validation study for the introduction of this rapid drug susceptibility test in Kinshasa, Democratic Republic of Congo. The double gyrA mutation 80Ala and 90Gly represented 57% of all fluoroquinolone mutations identified from MDR-TB patient sputum samples, as confirmed by DNA sequencing. This double mutation was previously found to be associated with susceptibility to fluoroquinolones, yet it leads to absent hybridization of a wild-type band in the MTBDRsl and is thus falsely scored as resistance. Our findings suggest that MTBDRsl results must be interpreted with caution when the interpretation is based solely on the absence of a wild-type band without confirmation by visualization of a mutant band. Performance of the MTBDRsl LPA might be improved by replacing the gyrA wild-type probes by additional probes specific for well-documented gyrA mutations that confer clinically relevant resistance.