Table 3. Optimization of primer concentrations and ratios in LAMP reaction.
| Loop Primer concentration | ||||||||
| 5.0μM | 4.2μM | 3.6μM | 2.8μM | |||||
| DNA concentration | BCG | H37Rv | BCG | H37Rv | BCG | H37Rv | BCG | H37Rv |
| 5pg | 21.0±1.3 | 82.1±5.9 | 23.8±0.6 | 95.3±2.6a | 24.8±0.6 | NA | 35.2±3 | NA |
| 500fg | 24±1.7 | NT | 24.9±1.2 | NT | 31.6±2.1 | NT | 39.9±10.1 | NT |
| 50fg | 27.1±0.9 | NT | 30.0±3.1 | NT | 36.1±4.0b | NT | NA | NT |
| NC | NA | NA | NA | NA | ||||
| Primer volume quantity | ||||||||
| 2.0μl | 2.25μl | 2.5μl | ||||||
| DNA concentration | BCG | H37Rv | BCG | H37Rv | BCG | H37Rv | ||
| 5pg | 37.6±1.6 | 117.3c | 32.8±4 | NA | 23.8±2.1 | 101.3±10.1d | ||
| 500fg | 41.9±6.2 | NT | 35.3±2.1 | NT | 31.9±7.0 | |||
| 50fg | 70±15.7e | NT | 45.1±9.0f | NT | 34.4±1.9 | |||
| NC | NA | NA | NA | |||||
LAMP reactions were performed six times (duplicate x 3). Results are shown in time (min.) to be positive presented as mean ± standard deviation. BCG: M. bovis BCG 172 Tokyo; H37Rv: M. tuberculosis H37Rv; NA: No amplification observed within 120 min of reaction; NT: Not tested; NC: Negative control (DDW); NA: No Amplification
a 2 of 6 reactions were positive
b 5 of 6 reactions were positive
c 1 of 6 reactions were positive
d 4 of 6 reactions were positive
e 3 of 6 reactions were positive
f 4 of 6 reactions were positive