Table 2. PCR assay validation results.
| Assaya | (sub)type | Slopeb | Eb | R2 b | LODc | Reproducibilityd | |||||
| Intrassay | Interassay | ||||||||||
| 500,000 | 5,000 | 50 | 500,000 | 5,000 | 50 | ||||||
| AMsw | A(H1N1) | −3.18 | 106% | 0.999 | 5.95 | 0.29 | 0.22 | 0.58 | 0.36 | 0.33 | 0.81 |
| A(H3N2) | −3.42 | 96% | 0.998 | 4.99 | 0.13 | 0.12 | 0.35 | 0.24 | 0.22 | 0.48 | |
| A(H1N1)v | −3.32 | 100% | 0.999 | 7.11 | 0.09 | 0.13 | 0.34 | 0.35 | 0.25 | 0.46 | |
| A(H5N1) | −3.68 | 87% | 0.999 | 6.79 | 0.27 | 0.14 | 0.46 | 0.39 | 0.18 | 0.40 | |
| AMsw+FCV | A(H1N1) | −3.55 | 91% | 0.999 | 12.81 | 0.19 | 0.13 | 0.59 | 0.20 | 0.44 | 0.61 |
| H1 | A(H1N1) | −3.40 | 97% | 0.998 | 2.81 | 0.10 | 0.11 | 0.21 | 0.14 | 0.21 | 0.36 |
| H3 | A(H3N2) | −3.70 | 86% | 0.997 | 4.92 | 0.14 | 0.21 | 0.46 | 0.25 | 0.29 | 0.54 |
| H1v * | A(H1N1)v | −3.14 | 108% | 0.987 | 14.33 | 0.12 | 0.27 | 0.67 | 0.22 | 0.29 | 0.52 |
| N1 | A(H1N1) | −3.39 | 97% | 0.998 | 6.49 | 0.18 | 0.16 | 0.51 | 0.31 | 0.16 | 0.41 |
| N2 | A(H3N2) | −3.46 | 95% | 0.999 | 6.79 | 0.18 | 0.10 | 0.28 | 0.16 | 0.09 | 0.54 |
| N1v * | A(H1N1)v | −3.30 | 101% | 0.999 | 9.76 | 0.32 | 0.21 | 0.55 | 0.29 | 0.32 | 0.64 |
| BM | B | −3.27 | 102% | 0.999 | 5.61 | 0.14 | 0.22 | 0.73 | 0.19 | 0.22 | 0.78 |
a
validation data was obtained on a Stratagene Mx3000 instrument for the assays indicated with an asterisk. All other assays were validated on a Stratagene Mx3005 instrument.
b
slope, efficiency (E) and correlation (R2) of standard curve; PCR efficiency was calculated as E = 10(−1/slope)−1.
c
Limit of detection (LOD) was calculated as 95% detection probability by probit analyses applying the SPSS 17.0 Statistics software.
d
reproducibility was calculated by examination of indicated plasmid copy numbers; intraassay: sixfold examination in a single run, interassay: twofold examination of double reactions plus inclusion of intraassay data; standard deviation of obtained Ct values are listed.