Table 5. Average quantification of the relative expression level, by RT-qPCR, of the genes that code for efflux pumps in M. tuberculosis in the two independent isoniazid exposure processes.
Relative expression level ± SD | ||||||
mmpl7 | Rv1258c | p55 | efpA | mmr | Rv2459 | |
Fully susceptible strains | ||||||
H37Rv INH(a)1 | 8.00±2.38 | 16.00±1.16 | 13.00±2.23 | 9.85±1.41 | 16.65±2.44 | 25.99±2.56 |
H37Rv INH(a)26 | 0.44±0.12 | 1.99±0.66 | 0.82±0.15 | 0.34±0.28 | 0.44±0.16 | 0.50±0.02 |
H37Rv INH(b)1 | 10.56±3.48 | 15.26±0.46 | 6.96±1.36 | 8.00±2.98 | 9.95±2.03 | 22.63±1.56 |
H37Rv INH(b)26 | 4.57±0.25 | 2.53±0.03 | 2.70±0.43 | 4.41±0.21 | 1.54±0.29 | 2.31±0.13 |
401/06 INH(a)1 | 34.30±1.24 | 22.63±0.47 | 18.38±1.94 | 16.00±1.32 | 24.25±2.34 | 9.19±0.11 |
401/06 INH(a)26 | 17.15±0.23 | 14.93±2.30 | 9.85±0.99 | 6.96±1.57 | 9.19±1.76 | 27.86±1.3 |
401/06 INH(b)1 | 1.53±0.29 | 1.77±0.75 | 3.47±1.16 | 6.50±1.30 | 1.47±0.07 | 0.20±0.05 |
401/06 INH(b)26 | 4.16±0.66 | 7.80±4.96 | 11.31±1.11 | 8.57±2.66 | 2.29±0.77 | 2.64±0.39 |
Rifampicin monoresistant strains | ||||||
267/05 INH(a)1 | 6.06±0.53 | 4.29±0.24 | 5.28±0.48 | 9.85±0.17 | 3.73±0.13 | 6.50±1.03 |
267/05 INH(a)26 | 115.56±59.44 | 53.85±2.64 | 110.80±66.17 | 341.05±66.43 | 59.06±53.66 | 32.00±1.98 |
267/05 INH(b)1 | 5.01±1.10 | 2.00±0.21 | 3.10±0.43 | 3.90±0.28 | 2.62±0.54 | 1.69±0.36 |
267/05 INH(b)26 | 17.15±1.24 | 3.04±0.29 | 12.13±0.67 | 22.63±3.09 | 3.89±0.57 | 0.64±0.09 |
359/03 INH(a)1 | 4.00±0.03 | 1.62±0.54 | 2.08±1.06 | 1.87±0.00 | 2.03±0.86 | 0.76±0.09 |
359/03 INH(a)26 | 13.96±1.36 | 9.01±2.17 | 8.53±2.86 | 10.66±2.07 | 11.46±4.9 | 32.00±2.01 |
359/03 INH(b)1 | 9.85±2.35 | 5.66±1.24 | 4.00±0.06 | 5.11±2.62 | 6.06±0.54 | 0.25±0.03 |
359/03 INH(b)26 | 25.99±1.33 | 29.86±3.29 | 29.86±2.56 | 2.30±0.83 | 10.56±1.36 | 2.30±0.12 |
Legend: (a)1: exposition process A, first passage; (b)1: exposition process B, first passage; (a)26: exposition process A, passage twenty six; (b)26: exposition process B, passage twenty six. The relative expression of the efflux pump genes was assessed by comparison of the relative quantity of the respective mRNA in the presence of isoniazid to the non-exposed strain. Each culture was assayed in triplicate using total RNA obtained from three independent cultures. A level of relative expression equal to 1 indicates that the expression level was identical to the strain that was being compared. Genes showing expression levels equal or above four, when compared to the non-exposed strain, were considered to be overexpressed and are shown in bold.