Detection of single HPV copies in SiHa cells by in situ polymerase chain reaction (in situ PCR) combined with immunoperoxidase and immunogold-silver staining (IGSS) techniques.
Detection of HPV by means of in situ hybridization techniques may often present problems if the number of HPV copies is too small, especially when using nonradioactive detection systems. A new way of amplifying extremely small amounts of virus DNA copies is the polymerase chain reaction (PCR), which is mostly used in vitro. In this study, we present a method for in situ PCR combined with immunogold-silver staining (IGSS) and immunoperoxidase (IMP) methods which allows the detection of single copies of HPV-DNA in SiHa cells. This method may have wide applications in routine diagnostic histopathology.
Zehbe
I
I
Department of Pathology, University Hospital, Uppsala, Sweden.
Hacker
G W
GW
Rylander
E
E
Sällström
J
J
Wilander
E
E
D001483:Base Sequence; D004279:DNA, Viral / Q000032:analysis* / Q000235:genetics; D005260:Female; D006801:Humans; D007124:Immunoenzyme Techniques; D007150:Immunohistochemistry / Q000379:methods; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D027383:Papillomaviridae / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods; D014407:Tumor Cells, Cultured; D002583:Uterine Cervical Neoplasms / Q000382:microbiology*
D004279:DNA, Viral; D009838:Oligodeoxyribonucleotides