Abstract
Detection of hepatitis B virus (HBV)-DNA in the liver of chronic infected patients is presently the most sensitive marker of viral replication and infectivity. In situ hybridization (ISH) allows the direct visualization of HBV infected liver cells and distribution of the viral sequences. This study was done to establish ISH and correlate the findings with conventional markers for HBV infection. Methods. Liver biopsies of 50 patients (28 ♂, 22 ♀) aged 0.5-20 years (mean 10.3) with various histological diagnoses were tested by 1SH. The HBV-DNA probe was labeled by nick translation with 35S-CTP to a specific activity of 3-5×108 cpm/μg DNA. Results. HBV-DNA/mRNA could be demonstrated in 38 patients, 12 were negative. Distribution of the grains was homogenous, inhomogenous with focal patches and focal. 33/38 children with HBeAg and 4/11 with anti-HBe were positive by ISH, 5/38 with HBcAg and 7/11 with anti-HBe remained negative. 17/23 HBsAg carriers with positive HBV-DNA/mRNA by ISH were positive for MBcAg in the liver and 31/36 had free HBV-DNA in Southern blot hybridization. Conclusions. Our results indicate that hepatitis B viral genomes can reliably be detected by in situ hybridization. Although there is a good correlation to other HBV markers (HBcAg, HBcAg and Southern blot hybridization) ISH may represent a more sensitive method to prove viral replication and infectivity.
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