Table 1.
qPCR Validation Data
| qPCR Assay | InsR-A | InsR-B | IGF-1R |
|---|---|---|---|
|
Amplification Efficiency Ave±SD, (n=10) |
100.71% ± 3.28% | 99.98% ± 2.32% | 100.49% ± 1.74% |
|
Slope Ave±SD, (n=10) |
−3.305 ± 0.077 | −3.322 ± 0.054 | −3.310 ± 0.041 |
|
Intercept Ave±SD, (n=10) |
36.97 ± 0.170 | 39.88 ± 0.376 | 37.34 ± 0.230 |
|
R squared Ave±SD, (n=10) |
0.996 ± 0.003 | 0.975 ± 0.011 | 0.998 ± 0.001 |
|
LOD copies/reaction (n=48) |
5 | 4 | 4 |
|
Intra-assay variation Ave±SD, (n=10) |
1.03% ± 0.86% | 0.71% ± 0.55% | 0.83% ± 0.83% |
|
Overall CV Ave±SD, (n=10) |
1.073% ± 0.959% | 3.052% ± 2.6% | 0.97% ± 0.52% |
|
Range of Detection copies/reaction |
5–106.4 | 4–106.4 | 4–107 |
|
Average Recovery Ave±SEM, (n=45) |
94.87%± 4.83% | 97.27% ± 5.54% | 98.67% ± 3.24% |
| Isoform Crossreactivity | 0.188%1 | No | NA |
|
Detection Rate Assay Specific |
93.6% (193/206) | 91.7% (189/206) | 91.8% (56/61) |
1
Only when InsR-B ≥ 104.4 copies/reaction.