HBV-DNA by in situ hybridization. A method to improve sensitivity on formalin-fixed, paraffin-embedded liver biopsies.
In situ hybridization of hepatitis B virus DNA by means of biotinylated probes is a rapid specific histochemical method but it is not very sensitive in formalin-fixed, paraffin-embedded material. The specificity and sensitivity of a new method using a digitonin solution and hydrolytic bath before cytohybridization in 19 formalin-fixed, paraffin-embedded liver biopsies was evaluated and a noticeable increase in staining intensivity observed. Digitonin and hydrochloric acid at low concentration increase the permeability and the diffusion of the bio-probe, improving significantly the sensitivity of reaction. Hepatocytes containing few copies of HBV-DNA were revealed by this method and microscopic visualization of specimens with numerous viral episomes was considerably improved. Therefore cytohybridization can be a specific and sensitive method, even for formalin-fixed, paraffin-embedded material, revealing active HBV replication even in HBcAg negative hepatocytes.
Infantolino
D
D
Servizio di Anatomia ed Istologia Patologica, Ospedale S. Giacomo Apostolo, Castelfranco Veneto, Italy.
Pinarello
A
A
Ceccato
R
R
Barbazza
R
R
D004279:DNA, Viral / Q000032:analysis*; D004072:Digitonin; D006509:Hepatitis B / Q000175:diagnosis*; D006515:Hepatitis B virus / Q000302:isolation & purification*; D006801:Humans; D008099:Liver / Q000473:pathology*; D009693:Nucleic Acid Hybridization
D004279:DNA, Viral; D004072:Digitonin