Fig. 1.

Fig. 2.

Fig. 3.

Fig. 4.

Fig. 5.

Fig. 6.

Fig. 7.

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 | rrs rrs WT | No | (S) |
| 2 | Mabs | rrs rrs WT | No | (S) |
| 3 | Mabs | rrs rrs WT | No | (S) |
| 4 | Mabs | rrs rrs WT | No | (S) |
| 5 | Mabs | rrs rrs WT | No | (S) |
| 6 | Mabs | rrs rrs WT | No | (S) |
| 7 | Mmas | rrs rrs WT | No | nt |
| 8 | Mmas | rrs rrs WT | No | (S) |
| 9 | Mabs | rrs MUT1 (A1408G) | Yes | (S) |
| 10 | Mmas | rrs rrs WT | No | (S) |
| 11 | Mb ol | rrs rrs WT | No | (S) |
| 12 | Mabs | rrs rrs WT | No | (S) |
j_pjm-2023-048_tab_007
| Strip 1 | – a clinical strain of Mycobacterium abscessus subs. abscessus (band characteristic for M. abscessus subsp. abscessus; SP4, SP5, SP6, SP9, SP10); no point mutation was detected; |
| Strips 2-6, 10, 12 - | –clinical strains of M. abscessus subs. abscessus (band characteristic for M. abscessus subsp. abscessus; SP4, SP5, SP6, SP9, SP10); point mutation erm(41)T28 referring to macrolide resistance; |
| Strips 7, 9 | – clinical strains of M. abscessus subs. massiliense (band characteristic for M. abscessus subsp. massiliense; SP4, SP5, SP8, SP9); point mutation erm(41)T28 no clinical implication in case of M. masiliense; |
| Strips 8 | – clinical strains of M. abscessus subs. abscessus (band characteristic for M. abscessus subsp. abscessus; SP4, SP5, SP6, SP9, SP10); point mutations were detected: erm(41)T28 and rrl MUT referring to macrolide resistance as well as point mutation A1408G referring to amikacin resistance; |
| Strip 11 | – clinical strains of M. abscessus subs. bolletii (band characteristic for M. abscessus subsp. bolletii; SP4, SP5, SP6, SP7, SP9, SP10), point mutation erm(41)T28 referring to macrolide resistance. |
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 | rrl MUT1 | A2058C |
| rrl MUT2 | A2058G | ||
| – | A2058T | ||
| rrl MUT3 | A2059C | ||
| rrl MUT4 | A2059T | ||
| Aminoglycosides | 1406–1409 | rrs MUT1 | 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 | erm(41) C28 rrlWT | No | (S) |
| 2 | Mabs | erm(41)T28 rrlWT | Yes | (R) |
| 3 | Mabs | erm(41)T28 rrlWT | Yes | (R) |
| 4 | Mabs | erm(41)T28 rrlWT | Yes | (R) |
| 5 | Mabs | erm(41)T28 rrlWT | Yes | (R) |
| 6 | Mmas | erm(41)T28 rrlWT | Yes | (R) |
| 7 | Mmas | erm(41)T28* rrlWT | No | nt |
| 8 | Mmas | erm(41)T28* rrlWT | No | (S) |
| 9 | Mabs | erm(41)T28 rrlMUT | Yes | (S) |
| 10 | Mb ol | erm(41)T28* rrlWT | No | (S) |
| 11 | Mabs | erm(41)T28 rrlWT | Yes | (R) |
| 12 | Mabs | erm(41)T28 rrlWT | Yes | (R) |