Premium
In situ hybridization detection of single‐copy human papillomavirus on isolated cells, using a catalyzed signal amplification system: Genpoint™
Author(s) -
Lizard Gérard,
DémaresPoulet MarieJosé,
Roignot Patrick,
Gambert Philippe
Publication year - 2001
Publication title -
diagnostic cytopathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.417
H-Index - 65
eISSN - 1097-0339
pISSN - 8755-1039
DOI - 10.1002/1097-0339(200102)24:2<112::aid-dc1020>3.0.co;2-6
Subject(s) - human papillomavirus , in situ , in situ hybridization , medicine , papillomaviridae , microbiology and biotechnology , genetics , gene , cervical cancer , biology , cancer , gene expression , physics , meteorology
The performance and drawbacks of GenPoint™, which is a catalyzed signal amplification system for immunohistochemistry, have been evaluated for its ability to reveal human papillomavirus (HPV) DNA detected by in situ hybridization with biotinylated DNA probes. For this aim, formalin‐fixed cell deposits from carcinoma cells of the uterine cervix, CaSki, SiHa, and HeLa, containing, respectively, 600 copies of HPV DNA type 16, 1–2 copies of HPV DNA type 16, and 10–50 copies of HPV DNA type 18, were used, and the GenPoint™ method (consisting of successive incubations with peroxidase‐conjugated streptavidin, biotinyl tyramide, and peroxidase‐conjugated streptavidin) was compared to immunoenzymatic revelation procedures involving either a one‐step reaction (streptavidin‐alkaline phosphatase or streptavidin‐peroxidase), or a three‐step reaction (anti‐biotin mouse monoclonal antibody, rabbit anti‐mouse antiserum, and mouse APAAP complex). In these conditions, after analysis with a bright‐field microscope, GenPoint™ appeared the most sensitive method of revelation, easily allowing detection of 1–2 copies of HPV DNA on isolated cells by in situ hybridization. Diagn. Cytopathol. 24:112–116, 2001. © 2001 Wiley‐Liss, Inc.