Polymerase Chain Reaction (PCR) In Situ Hybridization: Detection of Human Papillomavirus (HPV) DNA in SiHa Cell Monolayers View Full Text


Ontology type: schema:Chapter     


Chapter Info

DATE

1994

AUTHORS

Ingeborg Zehbe , Jan Sällström , Erik Wilander , Gerhard W. Hacker , Eva Rylander , Anders Strand , Anton-Helmut Graf

ABSTRACT

In situ hybridization (ISH) with labelled cDNA probes has become a valuable tool for the detection of human papillomavirus (HPV) as it allows direct correlation of virus infection and morphological diagnosis: This method, however, is limited to detect 10-50 DNA copies per cell which was demonstrated on HeLa cells. In the following study, we present a more sensitive in situ method, PCR in situ hybridization (PISH), able to detect a single HPV-DNA copy per cell. We developed a model with SiHa cells containing 1-2 HPV 16 copies per cell. SiHa cells stain negative with in situ hybridization. After amplifying the HPV-DNA in situ using PCR prior to the hybridization step, we were able to get positive results using either enzymatic detection methods or immunogold silver staining (IGSS). More... »

PAGES

297-306

References to SciGraph publications

Book

TITLE

Modern Methods in Analytical Morphology

ISBN

978-1-4613-6079-7
978-1-4615-2532-5

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/978-1-4615-2532-5_18

DOI

http://dx.doi.org/10.1007/978-1-4615-2532-5_18

DIMENSIONS

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