Table 1. The effect of isothermal temperature on the end point limit of detection of both the RT-LAMP (un-labelled internal primers) and RT-LAMP-LFD (labelled internal primers) reaction.
| RT-LAMP - temperature/°C | |||||||||||||
| RNA copies/µl | Mean Ct Value | 55 | 55.3 | 56.5 | 58.3 | 60.6 | 63.2 | 65.9 | 68.5 | 71 | 73.1 | 74.6 | 75.4 |
| 102 | 32.3 | + | + | + | + | + | + | + | + | − | − | − | − |
| 101 | 35.5 | + | + | + | + | + | + | + | − | − | − | − | − |
| 100 | +/− | + | + | +/− | +/− | +/− | +/− | − | +/− | − | − | − | − |
| 10−1 | – | + | − | − | − | − | − | − | − | − | − | − | − |
| Neg control | – | + | − | − | − | − | − | − | − | − | − | − | − |
| RT-LAMP-LFD - temperature/°C | |||||||||||||
| RNA copies/µl | Mean Ct Value | 55 | 55.3 | 56.5 | 58.3 | 60.6 | 63.2 | 65.9 | 68.5 | 71 | 73.1 | 74.6 | 75.4 |
| 102 | 32.3 | + | + | + | + | + | + | + | + | − | − | − | − |
| 101 | 35.5 | + | + | + | + | + | + | − | + | − | − | − | − |
| 100 | +/− | +/− | +/− | +/− | +/− | + | + | − | − | − | − | − | − |
| 10−1 | − | − | − | − | − | − | − | − | − | − | − | − | − |
| Neg control | − | − | − | − | − | − | − | − | − | − | − | − | − |
RNA standards were used to define the limit of detection, with the RT-qPCR Ct values being displayed for each RNA dilution. The RNA standards span the limit of detection of the RT-qPCR assay.+indicates reaction was positive by gel electrophoresis and PicoGreen fluorescence analysis (for un-labelled primers) or by LFD analysis (for labelled primers). +/− indicates variable positivity amongst quadruplicates analysed for a given RNA copy number.