J G E Laumen1,2, S S Manoharan-Basil1, E Verhoeven1,3, S Abdellati4, I De Baetselier4, T Crucitti5, B B Xavier2, S Chapelle2, C Lammens2, C Van Dijck1,2, S Malhotra-Kumar2, C Kenyon1,6. 1. Institute of Tropical Medicine, Department of Clinical Sciences, STI Unit, Antwerp, Belgium. 2. University of Antwerp, Laboratory of Medical Microbiology, Vaccine and Infectious Disease Institute, Antwerp, Belgium. 3. Pfizer, Puurs, Belgium. 4. Institute of Tropical Medicine, Department of Clinical Sciences, Clinical Reference Laboratory, Antwerp, Belgium. 5. Centre Pasteur du Cameroun, Yaounde, Cameroon. 6. Department of Medicine, University of Cape Town, Cape Town, South Africa.
Abstract
BACKGROUND: The prevalence of azithromycin resistance in Neisseria gonorrhoeae is increasing in numerous populations worldwide. OBJECTIVES: To characterize the genetic pathways leading to high-level azithromycin resistance. METHODS: A customized morbidostat was used to subject two N. gonorrhoeae reference strains (WHO-F and WHO-X) to dynamically sustained azithromycin pressure. We tracked stepwise evolution of resistance by whole genome sequencing. RESULTS: Within 26 days, all cultures evolved high-level azithromycin resistance. Typically, the first step towards resistance was found in transitory mutations in genes rplD, rplV and rpmH (encoding the ribosomal proteins L4, L22 and L34 respectively), followed by mutations in the MtrCDE-encoded efflux pump and the 23S rRNA gene. Low- to high-level resistance was associated with mutations in the ribosomal proteins and MtrCDE efflux pump. However, high-level resistance was consistently associated with mutations in the 23S ribosomal RNA, mainly the well-known A2059G and C2611T mutations, but also at position A2058G. CONCLUSIONS: This study enabled us to track previously reported mutations and identify novel mutations in ribosomal proteins (L4, L22 and L34) that may play a role in the genesis of azithromycin resistance in N. gonorrhoeae.
BACKGROUND: The prevalence of azithromycin resistance in Neisseria gonorrhoeae is increasing in numerous populations worldwide. OBJECTIVES: To characterize the genetic pathways leading to high-level azithromycin resistance. METHODS: A customized morbidostat was used to subject two N. gonorrhoeae reference strains (WHO-F and WHO-X) to dynamically sustained azithromycin pressure. We tracked stepwise evolution of resistance by whole genome sequencing. RESULTS: Within 26 days, all cultures evolved high-level azithromycin resistance. Typically, the first step towards resistance was found in transitory mutations in genes rplD, rplV and rpmH (encoding the ribosomal proteins L4, L22 and L34 respectively), followed by mutations in the MtrCDE-encoded efflux pump and the 23S rRNA gene. Low- to high-level resistance was associated with mutations in the ribosomal proteins and MtrCDE efflux pump. However, high-level resistance was consistently associated with mutations in the 23S ribosomal RNA, mainly the well-known A2059G and C2611T mutations, but also at position A2058G. CONCLUSIONS: This study enabled us to track previously reported mutations and identify novel mutations in ribosomal proteins (L4, L22 and L34) that may play a role in the genesis of azithromycin resistance in N. gonorrhoeae.
Authors: Jordan C Raisman; Michael A Fiore; Lucille Tomin; Joseph K O Adjei; Virginia X Aswad; Jonathan Chu; Christina J Domondon; Ben A Donahue; Claudia A Masciotti; Connor G McGrath; Jo Melita; Paul A Podbielski; Madelyn R Schreiner; Lauren J Trumpore; Peter C Wengert; Emalee A Wrightstone; André O Hudson; Crista B Wadsworth Journal: PLoS One Date: 2022-01-13 Impact factor: 3.240
Authors: Jolein G E Laumen; Christophe Van Dijck; Sheeba S Manoharan-Basil; Saïd Abdellati; Irith De Baetselier; Vicky Cuylaerts; Tessa De Block; Dorien Van den Bossche; Basil B Xavier; Surbhi Malhotra-Kumar; Chris Kenyon Journal: Front Microbiol Date: 2021-11-25 Impact factor: 5.640
Authors: Tessa de Block; Natalia González; Saïd Abdellati; Jolein Gyonne Elise Laumen; Christophe Van Dijck; Irith De Baetselier; Dorien Van den Bossche; Sheeba S Manoharan-Basil; Chris Kenyon Journal: Front Microbiol Date: 2022-03-30 Impact factor: 5.640