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Detection of supercoiled hepatitis B virus DNA and related forms by means of molecular hybridization to an oligonucleotide probe
Author(s) -
Lin Hsiang Ju,
Chung Hau Tim,
Lai Ching Lung,
Leong Susan,
Tam Oi Sin
Publication year - 1989
Publication title -
journal of medical virology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 121
eISSN - 1096-9071
pISSN - 0146-6615
DOI - 10.1002/jmv.1890290412
Subject(s) - dna supercoil , virology , dna , hepatitis b virus , oligonucleotide , microbiology and biotechnology , biology , hbsag , polymerase chain reaction , virus , hepatitis b virus pre beta , hybridization probe , hepatitis b virus dna polymerase , gene , genetics , dna replication
A novel assay for supercoiled and other fully double‐stranded forms of hepatitis B virus (HBV) DNA in blood is presented that utilizes molecular hybridisation to a radiophosphorous‐labeled oligonucleotide probe. The probe [5′‐d(ACGTGCA‐GAGGTGAAGCGA)] is complementary to the S(+)‐strand sequence furthest downstream, at the end of the gap. We examined blood specimens from 137 healthy HBsAg‐positive individuals, applying the probe to dots representing 2–3.5 ml serum or plasma. We found that supercoiled HBV is present in many HBV DNA‐positive blood specimens albeit in small quantities. Of the 104 specimens that were positive for HBV DNA of any form, 53 annealed to the probe. Serial specimens from the same subject taken over a period of months showed that the proportion of supercoil to other HBV DNA forms was variable. The presence of supercoil HBV DNA was not closely correlated with the level of serum HBV DNA polymerase. The supercoil is an HBV DNA form that can persist in the liver in the presence or absence of other replicative intermediates. This assay may enable further characterization of the status of HBV infection.