1.Viral DNA isolation
Human blood and plasma samples were spiked with 500 HBV copies per 200 µl sample. Viral DNA was isolated from 200 µl HBV infected human EDTA - blood using the Invisorb Spin Virus DNA Mini Kit and the respective kit from supplier Q. The extracted viral DNA was amplified on a Rotor-Gene™ 3000 using a HBV specific PCR. All test were done in triplicates.
2. Linear yields and reproducibility
Different viral DNA extractions were performed from HBV spiked plasma with a dilution series (50 copies/ prep – 50000 copies/ prep) using the Invisorb Spin Virus DNA Mini Kit. The figure shows the amplification results for 4 different concentrations of HBV in plasma. Each test was performed as duplicates.
Standard deviations were determined for HBV dilution series in the linear range of the appropriate downstream assay. For precise analysis the same downstream assay was used as for the determination of the linear range (preparation from given copy numbers was used as a standard).
Recoveries permit detection of less than 250 copies / ml of viral DNA by qPCR (inhouse) which shows that the Invisorb Spin Virus DNA Mini Kit provides 100% detection rate of HBV DNA from a 250-copy/ml (5 copies per PCR).