Molecular Identification of Strains within the Mycobacterium abscessus Complex and Determination of Resistance to Macrolides and Aminoglycosides
, , , y | 16 dic 2023
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Article Category: ORIGINAL PAPER
Publicado en línea: 16 dic 2023
Páginas: 491 - 506
Recibido: 15 sept 2023
Aceptado: 11 nov 2023
DOI: https://doi.org/10.33073/pjm-2023-048
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© 2023 Katarzyna Kania et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
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Results of susceptibility testing to antimycobacterial agents obtained by Sensititre™ RAPMYCO2 plates. Interpretation criteria following CLSI.
| MIC values (μg/ml) (Interpretation S/I/R) | ||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|
| No. | Species | AN | FOX | CIP | DOX | LZD | MXF | SXT | IMI | TGC | CLA after 5d | CLA after 14 d |
| 1 | Mabs | 16 (S) | 64 (I) | > 4 (R) | > 16 (R) | ≤ 1 (S) | 4 (R) | 4/76 (R) | 16 (I) | MIC = 1 | 0,06 (S) | 0,125 (S) |
| 2 | Mabs | 16 (S) | >128 (R) | > 4 (R) | > 16 (R) | 4 (S) | 2 (I) | > 8/152 (R) | 16 (I) | MIC = 0,5 | 0,06 (S) | > 16 (R) |
| 3 | Mabs | 16 (S) | 32 (I) | > 4 (R) | > 16 (R) | 4 (S) | > 8 (R) | 4/76 (R) | 16 (I) | MIC = 0,5 | 0,5 (S) | > 16 (R) |
| 4 | Mabs | 16 (S) | >128 (R) | > 4 (R) | > 16 (R) | 16 (I) | > 8 (R) | > 8/152 (R) | 16 (I) | MIC = 2 | 0,06 (S) | > 16 (R) |
| 5 | Mabs | 16 (S) | 64 (I) | > 4 (R) | > 16 (R) | 16 (I) | > 8 (R) | > 8/152 (R) | 16 (I) | MIC = 1 | 0,06 (S) | > 16 (R) |
| 6 | Mabs | 16 (S) | 64 (I) | > 4 (R) | > 16 (R) | 4 (S) | > 8 (R) | > 8/152 (R) | 16 (I) | MIC = 2 | 4 (I) | > 16 (R) |
| 7 | Mmas | - | - | - | - | - | - | - | - | - | - | - |
| 8 | Mmas | 16 (S) | >128 (R) | > 4 (R) | > 16 (R) | 16 (I) | 4 (R) | 4/76 (R) | 16 (I) | MIC = 0,25 | 0,06 (S) | 0,125 (S) |
| 9 | Mabs | 16 (S) | >128 (R) | > 4 (R) | > 16 (R) | 16 (I) | 4 (R) | 4/76 (R) | 16 (I) | MIC = 0,5 | 0,06 (S) | 0,125 (S) |
| 10 | Mmas | 16 (S) | 32 (I) | > 4 (R) | 1 (S) | 1 (S) | 2 (I) | 2/38 (S) | 16 (I) | MIC = 0,5 | 0,06 (S) | 0,06 (S) |
| 11 | Mbol | 4(S) | 32 (I) | > 4 (R) | > 16 (R) | 4 (S) | > 8 (R) | > 8/152 (R) | 16 (I) | MIC = 0.25 | 0.06 (S) | > 16 (R) |
| 12 | Mabs | 4 (S) | 33 (I) | > 4 (R) | > 16 (R) | 8 (S) | > 8 (R) | > 8/152 (R) | 16 (I) | MIC = 0.12 | 4 (I) | > 16 (R) |
Interpretation of susceptibility pattern of rapidly growing mycobacteria according to the CLSI recommendations.
| Antimycobacterial agents | MIC breakpoints (μg/ml) | ||
|---|---|---|---|
| S | I | R | |
| Amikacin | ≤ 16 | 32 | ≥ 64 |
| Cefoxitin | ≤ 16 | 32-64 | ≥ 128 |
| Doxycycline | ≤ 1 | 2-4 | ≥ 32 |
| Ciprofloxacin | ≤ 1 | 2 | ≥ 4 |
| Clarithromycin | ≤ 2 | 4 | ≥ 8 |
| Linezolid | ≤ 8 | 16 | ≥ 32 |
| Moxifloxacin | ≤ 1 | 2 | ≥ 4 |
| Trimthoprim/sulfamethoazole | ≤ 2/38 | – | ≥ 4/76 |
| Imipenem | ≤ 4 | 8-16 | ≥ 32 |
| Tigecycline |
– | – | – |
Results of molecular identification of resistance mechanisms to amikacin obtained by GenoType NTM in relation to phenotypic susceptibility testing results.
| No. of strain | Species | GenoType NTM-DR results | Phenotypic results of susceptibility to amikacin | |
|---|---|---|---|---|
| Type of bands (WT or mutation) | Molecular mechanisms of resistance (Yes/No) | |||
| 1 | Mabs | No | (S) | |
| 2 | Mabs | No | (S) | |
| 3 | Mabs | No | (S) | |
| 4 | Mabs | No | (S) | |
| 5 | Mabs | No | (S) | |
| 6 | Mabs | No | (S) | |
| 7 | Mmas | No | nt | |
| 8 | Mmas | No | (S) | |
| 9 | Mabs | Yes | (S) | |
| 10 | Mmas | No | (S) | |
| 11 | Mb ol | No | (S) | |
| 12 | Mabs | No | (S) | |
j.pjm-2023-048.tab.007
| Strip 1 | – a clinical strain of |
| Strips 2-6, 10, 12 - | –clinical strains of |
| Strips 7, 9 | – clinical strains of |
| Strips 8 | – clinical strains of |
| Strip 11 | – clinical strains of |
Interpretation of the mutation type conferring to the macrolides and aminoglicosides resistance according to the GenoType NTM-DR VER 1.0 test instructions.
| Discernible phenotypic pattern resistance | Target nucleic acid sequence | Nascent mutation band | Mutation |
|---|---|---|---|
| Macrolides | 2058–2059 | A2058C | |
| A2058G | |||
| – | A2058T | ||
| A2059C | |||
| A2059T | |||
| Aminoglycosides | 1406–1409 | A1408G | |
| – | T1406A | ||
| – | C1409T |
Results of susceptibility testing to amikacin and clarithromycin obtained by manually conducted microdilution method. Interpretation criteria following CLSI.
| No. | Species | AN | Interpretation |
CLA after 5 days of incubation | Interpretation |
CLA after 14 days of incubation | Interpretation |
|---|---|---|---|---|---|---|---|
| 1 | Mabs | 16 | (S) | 0.06 | (S) | 0,5 | (S) |
| 2 | Mabs | 8 | (S) | 0.06 | (S) | > 64 | (R) |
| 3 | Mabs | 16 | (S) | 0.06 | (S) | = 32 | (R) |
| 4 | Mabs | 16 | (S) | 0.06 | (S) | = 32 | (R) |
| 5 | Mabs | 16 | (S) | 0.06 | (S) | = 32 | (R) |
| 6 | Mabs | 16 | (S) | 0.06 | (S) | = 32 | (R) |
| 7 | Mmas | – | – | – | – | – | – |
| 8 | Mmas | 16 | (S) | 0.06 | (S) | = 0.125 | (S) |
| 9 | Mabs | 16 | (S) | 0.06 | (S) | = 0.125 | (S) |
| 10 | Mmas | 16 | (S) | 0.06 | (S) | = 0.06 | (S) |
| 11 | Mbol | 0.5 | (S) | 0.06 | (S) | > 64 | (R) |
| 12 | Mabs | 0.250 | (S) | 4 | (R) | = 16 | (R) |
Results of molecular identification of resistance mechanisms to clarithromycin by GenoType NTM in relation to phenotypic susceptibility testing results.
| No. of strain | Species | GenoType NTM-DR results | Phenotypic results of susceptibility to clarithromycin | |
|---|---|---|---|---|
| Type of bands (WT or mutation) | Molecular mechanisms of resistance (Yes/No) | |||
| 1 | Mabs | No | (S) | |
| 2 | Mabs | Yes | (R) | |
| 3 | Mabs | Yes | (R) | |
| 4 | Mabs | Yes | (R) | |
| 5 | Mabs | Yes | (R) | |
| 6 | Mmas | Yes | (R) | |
| 7 | Mmas | No | nt | |
| 8 | Mmas | No | (S) | |
| 9 | Mabs | Yes | (S) | |
| 10 | Mb ol | No | (S) | |
| 11 | Mabs | Yes | (R) | |
| 12 | Mabs | Yes | (R) | |