Table 8.
Contaminating DNA Q-PCR assays validated to ICH guidelines
| Host cell | Target gene | Sensitivity | Sensitivity |
|---|---|---|---|
| (Q-PCR) | Q-PCR | hybridisationb | |
| 293 | Adenovirus E1 | 500 fg–5 pg | 50 pg |
| E. coli | 23S RNA | 500 fga | 100 pg |
| HeLa | HPV-18 E7 | 5–50 pg | 50 pg |
| Primate | β-actin | 5–50 pg | 50 pg |
| Rodent | GAPDH | 500 fg–5 pg | 20 pg |
| (CHO) | |||
| Rodent | GAPDH | 500 fg–5 pg | 20 pg |
| (mouse) | |||
| Pichia | β-actin | 500 fg–5 pg | ND |
| Chicken | β-actin | 5 pg | ND |
| Porcine | PERV | 500 fg–5 pg | ND |
Abbreviations: ND: not determined; PERV: porcine endogenous retrovirus; GAPDH: glutaraldehyde-6-phosphate dehydrogenase.
a
Assay has background signals equivalent to that below 50 fg of E. coli DNA. These signals are due to residual DNA present in AmpliTaq DNA polymerase.
b
Slot-blot technique using 32P-radiolabeled probe (genomic DNA), hybridisation and autoradiography.