Detection of supercoiled hepatitis B virus DNA and related forms by means of molecular hybridization to an oligonucleotide probe

Journal Article · Fri Dec 01 00:00:00 EST 1989 · Journal of Medical Virology; (USA)

DOI:https://doi.org/10.1002/jmv.1890290412· OSTI ID:6937939

Lin, H J; Chung, H T; Lai, C L; Leong, S; Tam, O S ^[1]

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1. Univ. of Hong Kong (Hong Kong)

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(ACGTGCAGAGGTGAAGCGA)) 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.

• OSTI ID: 6937939
• Journal Information: Journal of Medical Virology; (USA), Vol. 29:4; ISSN 0146-6615
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
DNA POLYMERASES
BIOSYNTHESIS
HEPATITIS
BIOASSAY
ANTIGENS
DNA
ELECTROPHORESIS
MAN
OLIGONUCLEOTIDES
PHOSPHORUS ISOTOPES
SCINTILLATION COUNTING
TRACER TECHNIQUES
VIRUSES
ANIMALS
COUNTING TECHNIQUES
DIGESTIVE SYSTEM DISEASES
DISEASES
ENZYMES
ISOTOPE APPLICATIONS
ISOTOPES
MAMMALS
MICROORGANISMS
NUCLEIC ACIDS
NUCLEOTIDYLTRANSFERASES
ORGANIC COMPOUNDS
PARASITES
PHOSPHORUS-GROUP TRANSFERASES
POLYMERASES
PRIMATES
SYNTHESIS
TRANSFERASES
VERTEBRATES
550901* - Pathology- Tracer Techniques