Protocol for the use of self-reporting duplex mutation primers to detect PCR products in the diagnosis of HBV.

Quantitative measurements of serum hepatitis B virus (HBV) DNA are useful for tailoring of treatment schedules and the monitoring of HBV replication during therapy. We developed a novel fluorescence-based quantitative real-time PCR for quantitating HBV DNA based on the duplex mutation primers principle, in which signal is generated by melting a duplex mutation primer during renaturation. The duplex mutation primers are much more specific than double-stranded DNA dyes like SYBR Green I and, unlike other probes, do not require the double-labeled synthesis of fluorophore and quencher on the same molecule.