Table 1.
Overview of the relative SDs and genomic titer recoveries of different dPCR and qPCR methods
| Target | cdPCR (Stilla) |
ddPCR (Bio-Rad) |
ndPCR (Qiagen) |
mapdPCR (Thermo Fisher Scientific) |
qPCR (Bio-Rad) |
||||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| CV |
Recovery |
CV |
Recovery |
CV |
Recovery |
CV |
Recovery |
CV |
Recovery |
||
| (%) | (%) | (%) | (%) | (%) | (%) | (%) | (%) | (%) | (%) | ||
| AAV2 | SV40 | 10.4 | 95.6 | 14 | 78.3 | 9.6 | 98.9 | 6.2 | 62.7 | 33.6 | 103.1 |
| ITR | 9.6 | 126.5 | 13.5 | 109.3 | 9.2 | 136.7 | 6.8 | 88.5 | 43.3 | 1668.1 | |
| AAV5 | SV40 | 0.2 | 102.1 | 0.8 | 98 | 8.9 | 109.2 | 1.7 | 98.5 | 11.4 | 117.7 |
| ITR | 6.1 | 149.3 | 0.8 | 150.5 | 8.0 | 171.7 | 2.6 | 151.4 | 68.6 | 1450 | |
| AAV8 | SV40 | 8.6 | 116.1 | 3.2 | 97.6 | 8.6 | 133.3 | 6.8 | 96.6 | 31.5 | 130.7 |
| ITR | 0.5 | 146.0 | 2.9 | 131.5 | 5.9 | 176.5 | 8.4 | 132.3 | 43.5 | 1957.8 | |
VG titers of AAV2, AAV5, and AAV8 RSM (AMSBIO Europe) were measured by cdPCR, ndPCR, ddPCR, mapdPCR, and qPCR as a duplex assay targeting both the SV40 and ITR region on the AAV genome. The recovery values were calculated with respect to the specified VG titer. Experiments were performed as independent triplicate measurements. CV, coefficient of variation; SV40, simian virus 40, ITR, inverted terminal repeats.