Volume 28, Number 9—September 2022
Research
Rapid Adaptation of Established High-Throughput Molecular Testing Infrastructure for Monkeypox Virus Detection
Table 2
Software settings for run protocol for a dual-target MPXV assay rapidly adapted from established high-throughput molecular testing infrastructure*
Protocol setting |
Channel setting, use |
||||
---|---|---|---|---|---|
1, Not used |
2, Monkeypox |
3, Nonvariola |
4, Not used |
5, Internal control |
|
Relative fluorescence increase |
NA |
2 |
2 |
NA |
2 |
PCR cycling conditions | UNG incubation | Pre-PCR step | 1st measurement | 2nd measurement | Cooling |
No. cycles | Predefined | 1 | 5 | 45 | Predefined |
No. steps | Predefined | 3 | 2 | 2 | Predefined |
Temperature | Predefined | 55°C; 60°C; 65°C | 95°C; 55°C | 91°C; 58°C | Predefined |
Hold time | Predefined | 120 s; 360 s; 240 s | 5 s; 30 s | 5 s; 25 s | Predefined |
Data acquisition | Predefined | None | End of each cycle | End of each cycle | Predefined |
*Protocol was run on 400 µL swab samples. Protocol adapted for cobas omni Utility Channel (Roche Diagnostics, https://diagnostics.roche.com). Relative fluorescence increase thresholds were used for automated result calls. NA, not applicable; UNG, uracyl-N-glycosylase.