Abstract
The introduction of molecular biology techniques in the diagnostics of chronic hepatitis B virus infection proved HBV DNA to be the most sensitive marker of viral replication and infectivity. The aim of our study was to characterize the HBV DNA status in children with chronic hepatitis B with various molecular biology techniques in relation to conventional HBV markers.Methods: 206 sera of 172 and liver tissue of 108 children with chronic hepatitis B infection were investigated by dot blot-, Southern blot-, and in situ hybridization. In dot blot and Southern blot negative specimens polymerase chain reaction (PCR) was performed.Results: 111 of the 206 sera were positive for HBV DNA by dot blot hybridization. 78% of these patients had HBeAg and 7.7% anti-HBc. In 60 (92.3%) of the anti-HBc positive sera no HBV DNA could be delected. 83.9% of the dot blot negative children were HBV DNA positive by PCR comprising all HBeAg positive and 80% of the anti-HBc positive cases. HBV DNA studies in liver tissue by Southern blot revealed free HBV DNA in 74/103 (71.8%). HBV DNA integration was present in 2 children (1.9%). PCR in the Southern blot negative tissue specimens confirmed the presence of viral sequences in all HBeAg and in 63% of anti-HBc positive cases. In situ hybridization was applied to tissue sections of 63 children. HBV DNA was detected in 48 patients. 40 were HBeAg- and 6 anti-HBc positive. The distribution of HBV DNA in the tissue was classified as homogeneous, inhomogencous with focal patches and focal.Conclusions: In conclusion our results demonstrate that all HBeAg- and most of the anti-HBc positive children show viral sequences in serum and liver. Integration of HBV DNA into the liver cell genome can occur at an early stage of chronic disease but is not a very frequent event. Finally, in situ hybridization is a reliable method to delect HBV DNA in small amounts of liver tissue and to provide informations about the distribution of replicative viral sequences.
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