Twenty-six clinical isolates of Mycobacterium abscessus resistant to amikacin were identified. Most isolates were from patients with posttympanostomy tube placement otitis media or patients with cystic fibrosis who had received aminoglycoside therapy. Isolates were highly resistant (MICs ú1024 mg/mL) to amikacin, kanamycin, gentamicin, tobramycin, and neomycin (all 2-deoxystreptamine aminoglycosides) but not to streptomycin. Sequencing of their 16S ribosomal (r) RNA revealed that 16 (94%) of 17 had an AjG mutation at position 1408. In vitro -selected amikacinresistant mutants of M. abscessus and Mycobacterium chelonae had the same resistance phenotype, and 15 mutants all had the same AjG substitution at position 1408. Introducing an rRNA operon from Mycobacterium smegmatis with a mutated AjG at this position into a single functional allelic rRNA mutant of M. smegmatis produced the same aminoglycoside resistance phenotype. These studies demonstrate this 16S rRNA mutation is responsible for amikacin resistance in M. abscessus, which has only one copy of the rRNA operon.
Aminoglycosides have been a major component of therapyKanamycin and the closely related amikacin are used for for mycobacterial diseases since the recognition of the remarktreatment of multidrug resistant tuberculosis [13]. [19]. The genetic basis of mycobacterial is complex, with high-level resistance associated with point resistance to amikacin, kanamycin, and tobramycin has, until mutations involving the ribosomal protein S12 (rpsL gene) and recently [20,21], been unknown. the 16S ribosomal RNA (rRNA) gene (rrs) [6 -11]. The 16S Phenotypic studies have shown that spontaneous mutants rRNA gene region that is mutated involves a pseudoknot strucselected in the laboratory for resistance to amikacin, kanamyture that is believed to be stabilized by the presence of the cin, and tobramycin (2-deoxystreptamine aminoglycosides) are S12 protein [9]. Low-level resistance to streptomycin in M.not cross-resistant to streptomycin for either M. tuberculosis tuberculosis is yet to be explained on the basis of genetic [22] or the rapidly growing mycobacteria [23]. In addition, alterations [12] but is believed to result from mutations involvrecent studies with Mycobacterium smegmatis have suggested ing cell wall transport [7].that the 16S rRNA position 1408 (Escherichia coli numbering system) may be important for resistance to amikacin, tobramycin, and gentamicin [20,21]. In the present study, we inves-