New RAPMYCOI SensititreTM Antimicrobial Susceptibility Test for Atypical Rapidly Growing Mycobacteria (RGM)
Abstract
:1. Introduction
- (1)
- M. fortuitum (M. fortuitum, M. peregrinum, M. sengalense, M. porcinum, M. neworleansense, M. boenickei, M. houstonense, M. brisbanense, M. septicum, and M. setense),
- (2)
- M. chelonae/M. abscessus complex (M. chelonae, M. immunogenum, M. franklinii, M. salmoniphilum, M. abscessus subsp. abscessus, M. abscessus subsp. Massiliense, and M. abscessus subsp. bolletii),
- (3)
- M. smegmatis (M. smegmatis and M. goodii),
- (4)
- M. mucogenicum (M. mucogenicum, M. phocaicum, and M. aubagnense),
- (5)
- M. mageritense/M. wolinskyi,
- (6)
2. Materials and Methods
2.1. Bacterial Strains and Growth Conditions
2.2. Strain Identification
2.3. Molecular Determination of Antimicrobial Susceptibility to Macrolides and Aminoglycosides
- If the strain had a genotype in which C was at position 28 it meant that the tested strain was sensitive to macrolides.
- If the strain had a genotype in which T was at position 28 it meant that the tested strain was resistant to macrolides.
2.4. Phenotypic Determination of Antimicrobial Susceptibility Profile
3. Results
3.1. Mycobacterium abscessus subsp. abscessus
3.2. Mycobacterium abscessus subsp. massiliense
3.3. Mycobacterium chelonae
3.4. Mycobacterium mucogenicum
3.5. Mycobacterium fortuitum Complex
4. Discussion
- at least two NTM culture-positive sputa or one bronchial wash or lavage sample,
- a transbronchial or lung biopsy specimen with supporting mycobacterial histopathology and a positive NTM culture.
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Absence of Wild-Type Band | Analysed Nucleic Acid Positions | Mutation Bands Present | Mutation | Phenotypic Resistance |
---|---|---|---|---|
rrl WT | 2058–2059 | rrl MUT1 | A2058C | macrolides |
rrl MUT2 | A2058G | |||
A2058T | ||||
rrl MUT3 | A2059C | |||
rrl MUT4 | A2059G | |||
A2059T |
Absence of Wild-Type Band | Analysed Nucleic Acid Positions | Mutation Bands Present | Mutation | Phenotypic Resistance |
---|---|---|---|---|
rrs WT | 1406–1409 | rrs MUT1 | A1408G | aminoglycosides |
T1406A | ||||
C1409T |
Antimicrobial Agent | MIC (μg/mL) | Comment | ||
---|---|---|---|---|
S | I | R | ||
AMI | ≤16 | 32 | ≥64 | M. abscessus complex isolates with MIC of ≥64 μg/mL should be retested and/or the 16S rRNA gene sequenced to check for mutation |
FOX | ≤16 | 32–64 | ≥128 | |
CIP | ≤1 | 2 | ≥4 | Ciprofloxacin and levofloxacin are interchangeable, but both are less active than the newer B-methoxy-fluoroquinolones |
CLA | ≤2 | 4 | ≥8 | See text for information on the erm gene; clarithromycin and azithromycin are interchangeable clinically |
DOX | ≤1 | 2–4 | ≥8 | |
MIN | ≤1 | 2–4 | ≥8 | |
IMI | ≤4 | 8–16 | ≥32 | All isolates of M. fortuitum, M. smegmatis, and the M. mucogenicum group are presumed imipenem susceptible; imipenem MICs do not predict meropenem or ertapenem susceptibility |
LZD | ≤8 | 16 | ≥32 | |
MXF | ≤1 | 2 | ≥4 | |
TMP-SMX | ≤2/38 | ≥4/76 | MIC is 80% inhibition | |
TOB | ≤2 | 4 | ≥8 | Predominantly for M. chelonae; if MIC >4 μg/mL, the test should be repeated and/or the identification confirmed by rpoβ gene sequencing |
TGC | Insufficient data to establish breakpoints; only MIC should be reported |
M. abscessus subsp. abscessus n = 16 | M. abscessus subsp. massiliense n = 7 | M. chelonae n = 5 | M. mucogenicum n = 2 | M. fortuitum complex n = 30 | |
---|---|---|---|---|---|
ANTIBIOTIC AGENT | values in (%) | ||||
AMI | 100 (S) | 86 (S) 14 (R) | 100 (S) | 100 (S) | 100 (S) |
FOX | 100 (I) | 86 (I) 14 (S) | 80 (S) 20 (R) | 100 (S) | 67 (I) 33 (S) |
CIP | 81 (R) 19 (I) | 100 (R) | 100 (R) | 100 (S) | 97 (S) 3 (I) |
CLA | 75 (R) 25 (S) | 71 (S) 29 (R) | 100 (S) | 100 (S) | 77 (R) 23 (S) |
IMI | 100 (I) | 100 (I) | 80 (R) 20 (I) | 50 (S) 50 (I) | 63 (I) 27 (S) 10 (R) |
LZD | 75 (S) 25 (I) | 100 (S) | 100 (S) | 100 (S) | 93 (S) 7 (I) |
DOX | 100 (R) | 100 (R) | 100 (R) | 50 (S) 50 (R) | 56,6 (R) 43,3 (S) |
MIN | 100 (R) | 57 (S) 43 (R) | 100 (R) | 50 (S) 50 (I) | 56,6 (R) 43,3 (S) |
MXF | 81 (R) 19 (I) | 86 (R) 14 (I) | 80 (R) 20 (S) | 100 (S) | 100 (S) |
SXT | 100 (R) | 100 (R) | 100 (R) | 100 (S) | 70 (R) 30 (S) |
TOB | 100 (S) |
M. abscessus subsp. abscessus (n = 16) | |||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|
AMI | FOX | CIP | CLA | DOX | IMI | LZD | MIN | MXF | TGC | SXT | |
1 | 4 (S) | 32 (I) | 2 (I) | 0.5 (S) | >16 (R) | 16 (I) | 8 (S) | >8 (R) | 4 (R) | 0.5 | >8/152 (R) |
2 | 8 (S) | 32 (I) | 4 (R) | >16 (R) | 16 (R) | 16 (I) | ≤1 (S) | >8 (R) | 2 (I) | 0.5 | 8/152 (R) |
3 | 4 (S) | 32 (I) | 4 (R) | >16 (R) | >16 (R) | 16 (I) | 16 (I) | >8 (R) | 4 (R) | 0.5 | >8/152 (R) |
4 | 2 (S) | 32 (I) | 4 (R) | >16 (R) | >16 (R) | 16 (I) | 8 (S) | >8 (R) | 4 (R) | 0.5 | >8/152 (R) |
5 | 4 (S) | 32 (I) | 4 (R) | >16 (R) | >16 (R) | 16 (I) | 8 (S) | >8 (R) | 4 (R) | 0.12 | >8/152 (R) |
6 | 2 (S) | 32 (I) | >4 (R) | >16 (R) | >16 (R) | 8 (I) | 8 (S) | >8 (R) | 8(R) | 0.5 | >8/152 (R) |
7 | 4 (S) | 32 (I) | 4 (R) | >16 (R) | >16 (R) | 16 (I) | 2 (S) | >8 (R) | 4 (R) | 0.06 | 8/152 (R) |
8 | 4 (S) | 32 (I) | 2 (I) | 2 (S) | >16 (R) | 8 (I) | 8 (S) | >8 (R) | 4 (R) | 1 | 8/152 (R) |
9 | 8 (S) | 32 (I) | 4 (R) | 1 (S) | >16 (R) | 8 (I) | 4 (S) | >8 (R) | 4 (R) | 0.25 | >8/152 (R) |
10 | 4 (S) | 32 (I) | 4 (R) | >16 (R) | >16 (R) | 8 (I) | 8 (S) | >8 (R) | 4 (R) | 0.25 | >8/152 (R) |
11 | 4 (S) | 32 (I) | >4 (R) | >16 (R) | >16 (R) | 16 (I) | 16 (I) | >8 (R) | >8(R) | 1 | >8/152 (R) |
12 | 8 (S) | 32 (I) | >4 (R) | >16 (R) | >16 (R) | 16 (I) | 16 (I) | >8 (R) | >8 (R) | 1 | >8/152 (R) |
13 | 4 (S) | 32 (I) | 2 (I) | >16 (R) | >16 (R) | 16 (I) | 2 (S) | >8 (R) | 2 (I) | 0.25 | 4/76 (R) |
14 | 4 (S) | 32 (I) | 4 (R) | >16 (R) | >16 (R) | 16 (I) | 4 (S) | >8 (R) | 2 (I) | 0.25 | >8/152 (R) |
15 | 4 (S) | 64 (I) | 4 (R) | >16 (R) | >16 (R) | 16 (I) | 16 (I) | >8 (R) | 8 (R) | 0.5 | >8/152 (R) |
16 | 4 (S) | 32 (I) | >4 (R) | 0.12 (S) | >16 (R) | 16 (I) | 4 (S) | >8 (R) | 4 (R) | 0.25 | >8/152 (R) |
M. abscessus subsp. massiliense (n = 7) | |||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|
AMI | FOX | CIP | CLA | DOX | IMI | LZD | MIN | MXF | TGC | SXT | |
1 | 4 (S) | 32 (I) | 4 (R) | 0.25 (S) | >16 (R) | 16 (I) | 8 (S) | >8 (R) | 8 (R) | 1 | >8/152 (R) |
2 | >64 (R) | 16 (S) | 4 (R) | ≤0.06 (S) | >16 (R) | 16 (I) | 2 (S) | >8 (R) | 2 (I) | 0.12 | 8/152 (R) |
3 | 4 (S) | 32 (I) | 4(R) | 0.12 (S) | 8 (R) | 16 (I) | 8 (S) | 2 (S) | 4 (R) | 0.25 | >8/152 (R) |
4 | 8 (S) | 32 (I) | >4 (R) | 0.25 (S) | >16 (R) | 16 (I) | 8 (S) | >8 (R) | >8 (R) | 0.5 | >8/152 (R) |
5 | 4 (S) | 32 (I) | 4 (R) | >16 (R) | 16 (R) | 16 (I) | 8 (S) | 2 (S) | 8 (R) | 0.5 | 8/152 (R) |
6 | 4 (S) | 32 (I) | 4 (R) | >16 (R) | 16 (R) | 16 (I) | 8 (S) | 2 (S) | 8 (R) | 0.5 | 8/152 (R) |
7 | 8 (S) | 32 (I) | >4 (R) | 0.25 (S) | >16 (R) | 16 (I) | 8 (S) | 2 (S) | >8 (R) | 0.5 | >8/152 (R) |
M. chelone (n = 5) | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|
AMI | FOX | CIP | CLA | DOX | IMI | LZD | MIN | MXF | TGC | TOB | SXT | |
1 | 16 (S) | >128 (R) | 4(R) | 0.5 (S) | >16 (R) | 16 (I) | 4 (S) | >8 (R) | 1 (S) | 0.5 | 2 (S) | 8/152 (R) |
2 | 8 (S) | 64 (I) | 4(R) | ≤0.06 (S) | >16 (R) | 32 (R) | 4 (S) | >8 (R) | 4(R) | 0.25 | ≤1 (S) | 8/152 (R) |
3 | 4 (S) | 64 (I) | 4(R) | 0.25 (S) | >16 (R) | 32 (R) | 4 (S) | >8 (R) | 4(R) | 0.5 | ≤1 (S) | >8/152 (R) |
4 | 4 (S) | 64 (I) | 4(R) | 0.25 (S) | >16 (R) | 32 (R) | 4 (S) | >8 (R) | 4(R) | 0.5 | ≤1 (S) | 8/152 (R) |
5 | 8 (S) | 64 (I) | 4(R) | 0.25 (S) | >16 (R) | 64(R) | 4 (S) | >8 (R) | 4(R) | 0.25 | ≤1 (S) | >8/152 (R) |
M. mucogenicum (n = 2) | |||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|
AMI | FOX | CIP | CLA | DOX | IMI | LZD | MIN | MXF | TGC | SXT | |
1 | 2 (S) | 16 (S) | 0.25 (S) | 0.25 (S) | >16 (R) | 8 (I) | 2 (S) | >8 (R) | ≤0.25 (S) | 0.25 | 1/19 (S) |
2 | ≤1 (S) | 8 (S) | 0.5 (S) | 0.12 (S) | ≤0.12 (S) | 4 (S) | 2 (S) | ≤1 (S) | 0.5 (S) | 0.12 | 0.5/9.5 (S) |
M. fortuitum Complex (n = 30) | |||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|
AMI | FOX | CIP | CLA | DOX | IMI | LZD | MIN | MXF | TGC | SXT | |
1 | ≤1 (S) | 16 (S) | 0.5 (S) | 0.5 (S) | 0.5 (S) | ≤2 (S) | 4 (S) | ≤1 (S) | ≤0.25 (S) | 0.12 | 2/38 (S) |
2 | ≤1 (S) | 16 (S) | ≤0.12 (S) | 0.12 (S) | >16 (R) | ≤2 (S) | 2 (S) | >8 (R) | ≤0.25 (S) | 0.5 | 4/76 (R) |
3 | ≤1 (S) | 32 (I) | ≤0.12 (S) | >16 (R) | >16 (R) | 4 (S) | 2 (S) | >8 (R) | ≤0.25 (S) | 0.25 | 0.5/9.5 (S) |
4 | ≤1 (S) | 16 (S) | ≤0.12 (S) | 0.12 (S) | >16(R) | 4 (S) | ≤1 (S) | >8 (R) | ≤0.25 (S) | 0.25 | 1/19 (S) |
5 | ≤1 (S) | 16 (S) | ≤0.12 (S) | >16 (R) | ≤0.12 (S) | 8 (I) | ≤1 (S) | ≤1 (S) | ≤0.25 (S) | 0.25 | 2/38 (S) |
6 | ≤1 (S) | 32 (I) | ≤0.12 (S) | >16 (R) | >16 (R) | 8 (I) | 16 (I) | >8 (R) | ≤0.25 (S) | 0.5 | 8/152 (R) |
7 | ≤1 (S) | 16 (S) | 0.25 (S) | >16 (R) | 0.5 (S) | 4 (S) | ≤1 (S) | ≤1 (S) | ≤0.25 (S) | 0.25 | 1/19 (S) |
8 | ≤1 (S) | 16 (S) | ≤0.12 (S) | 0.25 (S) | >16(R) | 4 (S) | ≤1 (S) | >8 (R) | ≤0.25 (S) | 0.25 | 1/19 (S) |
9 | ≤1 (S) | 64 (I) | 2(I) | >16 (R) | 0.5 (S) | 64(R) | 8 (S) | >8 (R) | 1 (S) | 1 | >8/152 (R) |
10 | ≤1 (S) | 64 (I) | 0.25 (S) | >16 (R) | >16 (R) | 16 (I) | 8 (S) | >8 (R) | ≤0.25 (S) | 0.25 | >8/152 (R) |
11 | 2 (S) | 64 (I) | 0.25 (S) | >16 (R) | >16 (R) | 16 (I) | 8 (S) | >8 (R) | 0.5 (S) | 0.25 | 4/76 (R) |
12 | 4 (S) | 64 (I) | 0.25 (S) | >16 (R) | 0.25 (S) | 64 (R) | 4 (S) | ≤1 (S) | ≤0.25 (S) | 0.06 | 2/38 (S) |
13 | ≤1 (S) | 32 (I) | ≤0.12 (S) | >16 (R) | ≤0.12 (S) | 4 (S) | 4 (S) | ≤1 (S) | ≤0.25 (S) | 0.03 | 0.5/9.5 (S) |
14 | ≤1 (S) | 32 (I) | 0.25 (S) | >16 (R) | >16 (R) | 8 (I) | 8 (S) | >8 (R) | ≤0.25 (S) | 0.25 | 8/152 (R) |
15 | ≤1 (S) | 32 (I) | ≤0.12 (S) | >16 (R) | >16 (R) | 8 (I) | 8 (S) | >8 (R) | ≤0.25 (S) | 0.03 | 2/38 (S) |
16 | ≤1 (S) | 8 (S) | ≤0.12 (S) | >16 (R) | 8 (R) | 4 (S) | 2 (S) | ≤1 (S) | ≤0.25 (S) | 0.12 | 4/76 (R) |
17 | ≤1 (S) | 32 (I) | 0.25 (S) | >16 (R) | >16 (R) | 8 (I) | 8 (S) | >8 (R) | ≤0.25 (S) | 0.25 | >8/152(R) |
18 | ≤1 (S) | 32 (I) | 0.25 (S) | 16 (R) | >16 (R) | 8 (I) | 8 (S) | >8 (R) | ≤0.25 (S) | 0.03 | >8/152(R) |
19 | ≤1 (S) | 32 (I) | 0.25 (S) | 16 (R) | >16 (R) | 8 (I) | 8 (S) | >8 (R) | ≤0.25 (S) | 0.06 | >8/152(R) |
20 | ≤1 (S) | 32 (I) | ≤0.12 (S) | >16 (R) | 0.25 (S) | 16 (I) | 4 (S) | ≤1 (S) | ≤0.25 (S) | 0.06 | 4/76 (R) |
21 | ≤1 (S) | 32 (I) | ≤0.12 (S) | >16 (R) | >16(R) | 8 (I) | 2 (S) | >8(R) | ≤0.25 (S) | 0.12 | 4/76 (R) |
22 | ≤1 (S) | 32 (I) | 0.25 (S) | >16 (R) | 0.5 (S) | 8 (I) | 8 (S) | ≤1 (S) | ≤0.25 (S) | 0.5 | >8/152(R) |
23 | ≤1 (S) | 32 (I) | ≤0.12 (S) | >16 (R) | 0.12 (S) | 8 (I) | 4 (S) | ≤1 (S) | ≤0.25 (S) | 0.25 | 4/76 (R) |
24 | ≤1 (S) | 32 (I) | ≤0.12 (S) | >16 (R) | 0.25 (S) | 8 (I) | 4 (S) | ≤1 (S) | ≤0.25 (S) | 0.25 | 8/152 (R) |
25 | ≤1 (S) | 16 (S) | 0.5 (S) | 1 (S) | 0.25 (S) | 8 (I) | 8 (S) | ≤1 (S) | ≤0.25 (S) | 0.25 | 4/76 (R) |
26 | ≤1 (S) | 32 (I) | ≤0.12 (S) | >16 (R) | >16 (R) | 16 (I) | 16 (I) | >8 (R) | ≤0.25 (S) | 0.25 | 4/76 (R) |
27 | ≤1 (S) | 32 (I) | 0.5 (S) | 0.25 (S) | 0.25 (S) | 32R | 8 (S) | ≤1 (S) | ≤0.25 (S) | 0.25 | 4/76 (R) |
28 | ≤1 (S) | 16 (S) | ≤0.12 (S) | >16 (R) | 0.25 (S) | 16 (I) | 4 (S) | ≤1 (S) | ≤0.25 (S) | 0.25 | 8/152 (R) |
29 | ≤1 (S) | 16 (S) | ≤0.12 (S) | 0.12 (S) | 16 (R) | 8 (I) | 8 (S) | 8 (R) | ≤0.25 (S) | 0.12 | 4/76 (R) |
30 | ≤1 (S) | 32 (I) | ≤0.12 (S) | >16(R) | >16(R) | 16 (I) | 8 (S) | >8(R) | ≤0.25 (S) | 0.06 | >8/152 (R) |
Sensitivity to Clarithromycin on Days 3–5 of Incubation | Sensitivity to Clarithromycin on Day 14 of Incubation | Genetic Mechanisms | Subspecies of M. abscessus | Phenotypic Sensitivity to Macrolides |
---|---|---|---|---|
sensitive | sensitive | non-functional erm gene (41) | M. a. massiliense | sensitive to macrolides |
sensitive | resistant | functional erm gene (41) | M. a. abscessus M. a. bolletii | inducible resistance to macrolides |
resistant | resistant | 23S point mutation in rRNA | any of the above listed | high constitutive resistance to macrolides |
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Borek, A.; Zabost, A.; Głogowska, A.; Filipczak, D.; Augustynowicz-Kopeć, E. New RAPMYCOI SensititreTM Antimicrobial Susceptibility Test for Atypical Rapidly Growing Mycobacteria (RGM). Diagnostics 2022, 12, 1976. https://doi.org/10.3390/diagnostics12081976
Borek A, Zabost A, Głogowska A, Filipczak D, Augustynowicz-Kopeć E. New RAPMYCOI SensititreTM Antimicrobial Susceptibility Test for Atypical Rapidly Growing Mycobacteria (RGM). Diagnostics. 2022; 12(8):1976. https://doi.org/10.3390/diagnostics12081976
Chicago/Turabian StyleBorek, Anna, Anna Zabost, Agnieszka Głogowska, Dorota Filipczak, and Ewa Augustynowicz-Kopeć. 2022. "New RAPMYCOI SensititreTM Antimicrobial Susceptibility Test for Atypical Rapidly Growing Mycobacteria (RGM)" Diagnostics 12, no. 8: 1976. https://doi.org/10.3390/diagnostics12081976