Antisense oligodeoxynucleotide strategies have been employed in a variety of eukaryotic systems both to understand normal gene function and to block gene expression. Pharmacologically, ‘code blockers’ are ideal agents for antitumour and antimicrobial treatments because of their specific mode of action. Here we report the inhibition of duck hepatitis B virus (DHBV) by antisense oligodeoxynucleotides in primary duck hepatocyte cultures in vitro as well as in DHBV‐infected Pekin ducks in vivo. The most effective antisense oligodeoxynucleotide was directed against the 5′ region of the pre‐S gene and resulted in a complete inhibition of viral replication and gene expression in vitro and in vivo. These results demonstrate the application of antisense oligodeoxynucleotides in vivo and exemplify their potential as human antiviral therapeutics.
Factors determining the sensitivity and specificity of the spot-blot hybridization technique for the detection of hepatitis B virus DNA in serum were systematically investigated. Methods for pretreatment of serum samples, mode of application of the samples to the transfer membranes, blot treatment and hybridization conditions were all found to affect the sensitivity of the assay. The optimum hybridization procedure was found to be incubation of serum samples with salt, NaOH, formaldehyde and detergent, followed by spot application of the samples. This method specifically detected hepatitis B virus DNA in serum with a sensitivity 5 to 15 times higher than the presently used assay procedures.
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