Related Experiment Video
Updated: Sep 30, 2026
Detection of Low Copy Number Integrated Viral DNA Formed by In Vitro Hepatitis B Infection
Published on: November 7, 2018
In situ detection of mutated hepatitis B virus in microdissected, formalin-fixed liver tissues from patients with
N Kojima1, N Horiike, K Michitaka
1The Third Department of Internal Medicine, Ehime University School of Medicine, Japan.
Background/Aims:
Hepatitis B virus (HBV) quasispecies have been detected in patients with chronic hepatitis B. In order to elucidate the relationship between HBV mutation and liver cell necrosis in situ, we analyzed sublobule-sized specimens microdissected from formalin-fixed paraffin-embedded liver biopsy tissues taken from patients with chronic hepatitis B.
Methods:
The subjects were 20 patients with chronic hepatitis B. We extracted HBV-DNA from two sublobular regions of HBV-infected liver biopsy tissue, those with the most severe and the mildest hepatitis activity, demonstrated microscopically. The DNA coding sequence of the precore-core region of HBV was determined by amplifying the DNA by the polymerase chain reaction, followed by direct sequencing.
Results:
In all seven patients with minimal to mild hepatitis activity, but only 4 of 13 with moderate to severe activity, the amino acid sequence of the precore-core region of HBV obtained from the region with the most severe hepatitis activity showed over 99% homology with the corresponding sequence of HBV obtained from region with the mildest hepatitis activity (p<0.05).
Conclusion:
The differences between intrahepatic HBVs observed in patients with highly active hepatitis suggest that exacerbation of hepatitis in vivo is related to the appearance of variants in the precore-core region of HBV.
