Abstract
Introduction: A new line probe assay, the GenoType® NTM DR, has been developed for subspecies identification and detection of resistance to macrolides and aminoglycosides in clinical Mycobacterium abscessus isolates. We studied the performance of the test compared to DNA sequencing and phenotypic drug susceptibility testing (pDST). Methods: 48 clinical M. abscessus isolates collected between 2015 and 2016 were identified to the subspecies level and analysed for erm (41) genotype, rrl and rrs gene mutations by Sanger sequencing. Broth micro dilution was performed for pDST of Clarithromycin and Amikacin. The results were compared to those of the GenoType® NTM DR assay. Discordant results were further analysed by repeat pDST and whole genome sequencing (WGS). Results: 35 isolates were identified as M. abscessus subsp. abscessus , 6 as M. abscessus subsp. bolletii , and 7 as M. abscessus subsp. massiliense based on rpoB sequences. Concordance of GenoType® NTM DR results with Sanger sequencing was 92% for subspecies identification and 100% for erm (41), rrl, and rrs genotypes, respectively. GenoType® NTM DR and pDST results matched in 98% for Clarithromycin resistance and in 96% for Amikacin resistance when repeat pDST results were taken into account. Conclusion: The new GenoType® NTM DR assay is a valuable test for subspecies identification of M. abscessus isolates and detection of defined mutations conferring resistance to Amikacin and Clarithromycin. Discrepancies between the line probe assay and pDST mainly relate to variations in phenotypic test results.
Highlights
A new line probe assay, the GenoType® NTM DR, has been developed for subspecies identification and detection of resistance to macrolides and aminoglycosides in clinical Mycobacterium abscessus isolates
M. abscessus subsp. massiliense has a dysfunctional erm(41) gene due to a 2 bp deletion of nucleotides 64–65 and a 274 bp deletion of nucleotides 159–432 leading to a macrolide susceptible phenotype [9]
All four isolates displayed Clarithromycin susceptibility patterns compatible with M. abscessus subsp. massiliense and an erm(41) sequence showing the characteristic deletions of nucleotides 64–65 and 159–432
Summary
A new line probe assay, the GenoType® NTM DR, has been developed for subspecies identification and detection of resistance to macrolides and aminoglycosides in clinical Mycobacterium abscessus isolates. The primary innate mechanism of macrolide resistance in M. abscessus is the inducible expression of an erythromycin ribosomal methylase, Erm(41). Abscessus results in inducible resistance (T28) or phenotypic susceptibility (C28), respectively [10]. Massiliense has a dysfunctional erm(41) gene due to a 2 bp deletion of nucleotides 64–65 and a 274 bp deletion of nucleotides 159–432 leading to a macrolide susceptible phenotype [9]. Acquired high-level resistance to macrolides is due to point mutations at positions 2058 and 2059 in the 23S rRNA (rrl) gene [11]. Acquired high-level resistance to aminoglycosides is conferred by point mutations at
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Disclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.