Table 1.
Assay statistics and reproducibility
|
A. Fluorescent plate scanner and whole-well signal integration | |||||||
|---|---|---|---|---|---|---|---|
| |
|
Sensitivity (LOD) units/ml |
Variability intra-assay COV (%) |
Variability inter-assay COV (%) | |||
| Assay1 | Assay 2 | Assay 3 | Assay 4 | ||||
| IFNα |
VSV-DsRed2 |
1.51 +/− 0.12 |
0.9 |
2.9 |
0.7 |
1.7 |
2.4 |
| VSV-ZsGreen |
2.07 +/− 0.94 |
1.8 |
8.6 |
5.5 |
3.3 |
16.0 |
|
| IFNβ |
VSV-DsRed2 |
1.71 +/− 0.27 |
1.4 |
1.1 |
0.6 |
1.0 |
5.1 |
| VSV-ZsGreen |
2.57 +/− 0.52 |
1.9 |
4.1 |
2.2 |
3.2 |
7.9 |
|
|
B. Fluorescence/absorbance plate reader | |||||||
| |
|
Sensitivity (LOD) units/ml |
Variability intra-assay COV (%) |
Variability inter-assay COV (%) |
|||
| |
|
Assay 1 |
Assay 2 |
Assay 3 |
Assay 4 |
|
|
| |
VSV-DsRed2 |
1.29 +/− 0.34 |
4.0 |
1.7 |
1.9 |
5.7 |
5.7 |
| IFNα |
VSV-ZsGreen |
0.96 +/−0.18 |
1.2 |
1.1 |
1.7 |
2.5 |
3.6 |
| |
Crystal violet |
4.09 +/− 0.43 |
17.5 |
14.0 |
7.3 |
8.6 |
21.9 |
| |
VSV-DsRed2 |
2.71 +/− 0.29 |
3.0 |
1.8 |
1.7 |
3.6 |
6.4 |
| IFNβ |
VSV-ZsGreen |
1.87 +/− 0.11 |
2.3 |
3.1 |
0.7 |
1.8 |
4.4 |
| Crystal violet | 8.44 +/− 2.28 | 20.1 | 6.6 | 4.3 | 7.3 | 18.5 | |
Viral replication reporters and two methods of signal measurement were tested for the functional antiviral assays and calculated IC50 values statistically compared with the traditional crystal violet antiviral assay. Coefficients of variance were calculated with a minimum of four replicates. +/− values are 95% confidence intervals over all samples.