title: A quantitative competitive PCR assay for the covalently closed circular form of the duck hepatitis B virus.
abstract: A crucial step in the establishment and maintenance of a hepadnavirus infection is the formation of a pool of covalently closed circular viral genomes in the nucleus. Changes in the size of this pool occur when an infection is established, when acute infections are resolved, and when chronic infections are treated with antiviral drugs. However, the lack of a quantitative assay for the cccDNA form of the virus has hampered study of the biology of this replication intermediate. In response to this need we have devised a sensitive and accurate competitive PCR assay that is highly selective for the cccDNA form of the duck hepatitis B virus. Since only small amounts of DNA are needed for the assay, cccDNA pool sizes can be monitored in live animals using DNA derived from needle biopsies of infected liver.
authors.affiliation: Department of Medical Microbiology and Immunology, and Glaxo Wellcome-Heritage Research Institute, 622 HMRC, University of Alberta, AB., T6G 2S2, Edmonton, Canada. bill.addison@ualberta.ca
authors.forename: W R
authors.initials: WR
authors.lastname: Addison
authors.forename: W W
authors.initials: WW
authors.lastname: Wong
authors.forename: K P
authors.initials: KP
authors.lastname: Fischer
authors.forename: D L
authors.initials: DL
authors.lastname: Tyrrell
mesh_terms: D000818:Animals; D001706:Biopsy; D015139:Blotting, Southern; D004270:DNA, Circular / Q000032:analysis*; D004279:DNA, Viral / Q000032:analysis*; D004372:Ducks; D018347:Hepadnaviridae Infections / Q000821:virology*; D015611:Hepatitis B Virus, Duck / Q000235:genetics / Q000302:isolation & purification*; D008099:Liver / Q000821:virology; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D004270:DNA, Circular; D004279:DNA, Viral