Broth microdilution protocol for determining antimicrobial susceptibility of Legionella pneumophila to clinically relevant antimicrobials
Research output: Contribution to journal › Research article › Contributed › peer-review
Contributors
- Institute of Medical Microbiology and Virology
- Cardiff University
- University of Oxford
- NHS Greater Glasgow and Clyde
- University of Lyon
- Stavanger University Hospital
- University of Ljubljana
- Ente Ospedaliero Cantonale (EOC)
- Nabriva Therapeutics
- Vrije Universiteit Brussel
- Instituto de Salud Carlos III
- Cardiff & Vale University Health Board
- University Hospital Carl Gustav Carus Dresden
- Ben-Gurion University of the Negev
- British Society of Antimicrobial Chemotherapy
- National Health Service (NHS) England
Abstract
Currently there is no detailed, internationally agreed protocol defined to evaluate antimicrobial susceptibility testing (AST) for Legionella pneumophila (required to establish epidemiological cut-off value or "ECOFF" boundaries); therefore, antimicrobial resistance in these isolates cannot be defined. AST methods utilising media containing activated charcoal as an ingredient, to enable Legionella growth, are unreliable as noted in an internationally authored opinion paper and a new gold standard is required. Here we define a detailed protocol for broth microdilution (BMD) using defined cell culture collection-deposited control reference strains (Philadelphia-1 and Knoxville-1) as well as two accessible reference strains with moderately (lpeAB-carrying) and markedly (23S rRNA mutation-carrying) elevated azithromycin minimum inhibitory concentration (MIC). The defined protocol enables up to eight L. pneumophila strains to be set up on a single 96-well plate per antimicrobial tested. Initial ranges to routinely capture an MIC for these reference strains using clinically relevant antimicrobials azithromycin (0.01-0.25 mg/L), levofloxacin (0.008-0.03 mg/L), lefamulin (0.01-2 mg/L), rifampicin (0.0002-0.0008 mg/L) and doxycycline (0.25-16 mg/L) following incubation for 48 h at 37 °C in a shaking incubator have been empirically determined. Establishment of this internationally agreed protocol sets the scene for the next step: validation and comparison of antimicrobial ranges between international Legionella reference laboratories to establish putative resistance cut-off thresholds for these clinically relevant antimicrobials.
Details
| Original language | English |
|---|---|
| Article number | 107071 |
| Pages (from-to) | 107071 |
| Journal | Journal of microbiological methods |
| Volume | 228 |
| Early online date | 18 Dec 2024 |
| Publication status | Published - Jan 2025 |
| Peer-reviewed | Yes |
External IDs
| Scopus | 85212842011 |
|---|
Keywords
Keywords
- Antimicrobial resistance, Lefamulin, Legionella pneumophila, Macrolides, Susceptibility