In situ detection of mutated hepatitis B virus in microdissected, formalin-fixed liver tissues from patients with chronic hepatitis B.
OBJECTIVE
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).


CONCLUSIONS
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.
Kojima
N
N
The Third Department of Internal Medicine, Ehime University School of Medicine, Japan.
Horiike
N
N
Michitaka
K
K
Onji
M
M
D000293:Adolescent; D000328:Adult; D004279:DNA, Viral / Q000032:analysis*; D005260:Female; D005557:Formaldehyde; D006515:Hepatitis B virus / Q000235:genetics / Q000302:isolation & purification*; D019694:Hepatitis B, Chronic / Q000473:pathology / Q000821:virology*; D006801:Humans; D008099:Liver / Q000473:pathology / Q000821:virology*; D008297:Male; D008875:Middle Aged; D009154:Mutation; D016133:Polymerase Chain Reaction / Q000379:methods; D016707:Tissue Fixation
D004279:DNA, Viral; D005557:Formaldehyde