✦ LIBER ✦

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

✍ Scribed by W. J. Nicholson; S. H. Black; P. Simmonds; C-W. Chung; D. Aw; J. F. Peutherer

Publisher: John Wiley and Sons
Year: 1992
Tongue: English
Weight: 902 KB
Volume: 36
Category: Article
ISSN: 0146-6615
DOI: 10.1002/jmv.1890360105

No coin nor oath required. For personal study only.

✦ Synopsis

Abstract

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°/o 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.

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