Table 3.
Assay methodology.
| Publication | Growth calculation | Assay type | MOI | Concentration | Volume per assay (ml) | Media added pervolume of blood | Incubation time (h) | Replicates | Assay controls |
|---|---|---|---|---|---|---|---|---|---|
| Cheon et al. (16) | Δlog10CFU = log10(final) – log10(initial) | MGIT | NR | 10,000 CFU/ml (100,000 RLU/ml) | 0.6 | 1:1 RPMI + glutamine + 25 mM HEPES | 96 | 2 | Simultaneous direct mycobacterial inoculation of MGIT tube |
| Hoft et al. (17) | Mycobacterial inhibition index = (RLU at pre-BCG day 3 or day 4/RLU at pre-BCG day 0)/(Post-BCG day 3 or day 4 RLU/post-BCG day 0 RLU) | BCG-lux | NR | 10,000 CFU/ml (100,000 RLU/ml) | 1 | 1:2 RPMI | 96 | 3 | None reported |
| Kampmann et al. (18) | Growth ratio = RLU at T96/RLU at T0 | BCG-lux | NR | 1,000,000 CFU/ml (10,000,000 RLU/ml) | 1 | 1:1 RPMI | 96 | 3 | None reported |
| Fletcher et al. (19) | Δlog10 CFU per day = log((CFU of sample at T96/CFU of control at T96)/4) | MGIT | NR | 150 CFU in 600 μl | 0.6 | 1:1 RPMI | 96 | 2 | Simultaneous direct mycobacterial inoculation of MGIT tube (duplicate) |
| Martineau et al. (20) | Luminescence ratio = RLU at T24 or T96/RLU at T0 | BCG-lux | 1 | 300,000 CFU/ml | 1 | 1:1 RPMI + 2 mM glutamine + 25 mM HEPES | 96 | 3 | None reported |
| Eisen et al. (8) | (RLU at T96 – RLU at T0)/RLU of culture broth | BCG-lux | 30 | 10,000 CFU/ml (100,000 RLU/ml), 200 ul blood in each of quadruplet tests | 1 | 1:1 RPMI + 1% HEPES | 72 | 4 | Supplemented 7H9 broth; plasma |
| Kampmann et al. (21) | Growth ratio = RLU at T96/RLU at T0 | BCG-lux | NR | 1,000,000 CFU/ml (10,000,000 RLU/ml) | 1 | 1:1 RPMI | 96 | 3 | None reported |
| Tena et al. (22) | Growth ratio = RLU at T96/RLU at T0 | BCG-lux | NR | 1,000,000 CFU/ml (10,000,000 RLU/ml) | 1 | 1:1 RPMI | 96 | 3 | None reported |
| Kampmann et al. (11) | Growth ratio = (RLU at T96 – RLU at T0)/(RLU at T0) | BCG-lux | NR | 10,000 CFU/ml (100,000 RLU/ml) | 1 | 1:1 RPMI + 1% L-glutamine and heparin | 96 | 3 | Plasma |
| Martineau et al. (9) | Luminescence ratio = RLU at T96/RLU at T0 | BCG-lux | 1 | 300,000 CFU/ml | 1 | 1:1 RPMI + 2 mM glutamine + 25 mM HEPES | 96 | 3 | None reported |
| Baguma et al. (28) | Δlog10 CFU = log10(final) – log10(initial) | MGIT | NR | 8,500 – 2,4000 CFU/ml | 0.6 | 1:1 RPMI | 96 | 2 | Simultaneous direct mycobacterial inoculation of MGIT tube |
| O’Shea et al. (7) | Growth ratio = log10(CFU of sample/CFU of control) | MGIT | NR | 150 CFU/600 μl | 0.6 | 1:1 RPMI containing 10% pooled human serum + 2 mM L-glutamine and 25 mM HEPES | 96 | 2 | Simultaneous direct mycobacterial inoculation of MGIT tube (duplicate) |
| Wallis et al. (29) | Δlog10CFU = log10(final) – log10(initial) | MGIT | NR | 10,000 CFU/ml (100,000 RLU/ml) | 0.6 | 1:1 tissue culture medium |
72 | 2/1* | Simultaneous direct mycobacterial inoculation of MGIT tube |
| Nicol et al. (30) | Growth ratio = RLU at T96/RLU at T0 | BCG-lux | NR | 1,000,000 CFU/ml (10,000,000 RLU/ml) | 1 | 1:1 RPMI | 96 | 3 | None reported |
| O’Shea et al. (31) | Growth ratio = log10(CFU of sample/CFU of control) | MGIT | NR | 150 CFU/600 μl | 0.6 | 1:1 RPMI containing 10% pooled human serum + 2 mM L-glutamine and 25 mM HEPES | 96 | 2 | Simultaneous direct mycobacterial inoculation of MGIT tube (duplicate) |
Note MOI indicates the multiplicity of infection stated as the number of monocytes estimated to be present in the assay per colony forming unit of mycobacteria. RLU, relative light units; GI, growth index; CFU, colony forming units; BCG, bacille Calmette-Guerrin; MOI, Multiplicity of Infection, mycobacteria per macrophage; *Duplicate in Brazil, single in USA.