Comparison of hepatitis B virus subtyping of d/y determinants by radioimmunoprecipitation assay and the polymerase chain reaction

W J Nicholson, S H Black, P Simmonds, C W Chung, D Aw, J F Peutherer

Research output: Contribution to journalArticlepeer-review

Abstract / Description of output

Using a double polymerase chain reaction a method was devised for detecting and subtyping hepatitis B virus DNA in serum samples. Primers from the S-gene were selected from the sequence analyses of five HBV HBsAg subtypes, to amplify HBV DNA and subtype for y specific DNA. Thirty-eight samples were subtyped for d and y determinants by radioimmunoprecipitation assay (RIPA) and the polymerase chain reaction (PCR). Subtyping by PCR and RIPA was in agreement in 100% of subtype y samples and 83.3% of subtype d, giving an overall correlation of 92.1%. As a third comparison, 12 amplified samples were digested by the restriction enzyme Sau 3A, which differentiates between subtypes y and d. The digest results agreed with PCR in 83.3% of the samples. In addition, we compared our standard phenol/chloroform extraction against a rapid one step method. The phenol/chloroform stage was found to be essential for the removal of nucleases and polymerase inhibitors present in sera.
Original languageEnglish
Pages (from-to)21-7
Number of pages7
JournalJournal of Medical Virology
Volume36
Issue number1
DOIs
Publication statusPublished - Jan 1992

Keywords / Materials (for Non-textual outputs)

  • Base Sequence
  • Consensus Sequence
  • DNA, Viral
  • Hepatitis B Surface Antigens
  • Hepatitis B virus
  • Humans
  • Molecular Sequence Data
  • Polymerase Chain Reaction
  • Polymorphism, Restriction Fragment Length
  • Radioimmunoassay
  • Sequence Homology, Nucleic Acid

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