A rapid microtiter assay for duck hepatitis virus reverse transcriptase

The duck hepatitis B virus (DHBV) genome contains a pol gene that codes for the viral polymerase protein. This enzyme, which is essential for the replication of the virus, has multiple activities including an RNA directed DNA polymerase or reverse transcriptase (RT) activity, an RNase H activity, and a DNA-directed DNA polymerase activity. The assay described in this chapter is designed to measure the ability of test compounds to inhibit the RNA-directed DNA polymerase activity of the DHBV pol gene product. The assay is based on research performed in the laboratory of Dr. Christoph Seeger (1) and on the observation that the e stem loop structure in the pregenomic RNA is required for initiation of DNA synthesis (2-4).