DNA extraction from the sample using commercial kits.
Real-time PCR using primers and SYBR® Green I No ROX Intercalators for the quantification of HBV DNA, which were designed from the sequence of the non-coding 5'NCR region of the hepatitis C virus, according to Carman, W. et al. 1990. For the quantification, we constructed a standard curve starting from the DNA extracted from the quantified serum of a positive HBV patient (positive control). DNA amplification is detected as an increase in fluorescence. A Step One thermal cycler from Applied Biosystems is used for the realization of the technique.
Positive control: sera from HBV DNA positive patients and with known viral load.