Detection of varicella-zoster virus DNA in cultured cells and tissue specimens by in situ hybridization using biotin-labeled probes.
A method of in situ hybridization with biotinylated probes is described for specific detection of varicella-zoster virus (VZV) DNA in infected cell cultures and in human biopsied or autopsied materials. From molecularly cloned EcoRI-fragments of VZV DNA, we selected as probes for in situ hybridization three fragments (B, H, and K) which had any detectable homology neither with human cell DNA nor with DNAs of VZV-related viruses (herpes simplex virus type 1, type 2 and human cytomegalovirus). The procedure of in situ hybridization was based on that of Brigati et al. (Virology 126, 32-50, 1983), but following modifications were made. 1) The concentration of probe DNA was lowered to 100 ng per ml. 2) Streptavidin-biotin system was used for detection of hybridization. 3) Acid phosphatase was used to generate color development discernible from melanin present in skin. 4) Before hybridization, deparaffinized sections were treated with trypsin by the procedure routinely employed for detection of viral antigens in paraffin-embedded sections.
Hondo
R
R
Department of Pathology, University of Tokyo, Japan.
Yogo
Y
Y
Kurata
T
T
Aoyama
Y
Y
D002478:Cells, Cultured; D003001:Cloning, Molecular; D004279:DNA, Viral / Q000032:analysis*; D005455:Fluorescent Antibody Technique; D014645:Herpesvirus 3, Human / Q000032:analysis*; D006801:Humans; D009693:Nucleic Acid Hybridization
D004279:DNA, Viral