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Updated: Aug 6, 2026
RNAscope for In situ Detection of Transcriptionally Active Human Papillomavirus in Head and Neck Squamous Cell Carcinoma
Published on: March 11, 2014
Detection of oncogenic HPV and identification of 72Arg polymorphic p53 by in situ PCR for clinical routine purposes
Massimo Realacci1, Giulietta Anna Perrone, Patrizio Sale
1Department of Experimental Medicine and Pathology, University La Sapienza, Rome, Italy.
Background:
Ano-genital carcinoma is a polyfactorial and polygenic disease. Certain strains of human papillomavirus (HPV) have been detected in a high percentage of patients. It has been suggested that p53 polymorphisms may be relevant for the interaction with viral proteins that inactivates p53.
Materials And Methods:
Patients were selected on the basis of HPV infection, clinical history, positive PAP test and type of lesion. In situ PCR was performed on smear samples, in four steps: a) preparation on clean biobond-treated slides, b) permeabilisation and digestion; c) in situ PCR amplification; d) in situ hybridisation with a fluorescent probe.
Results:
In situ PCR analysis of the smears confirmed the results obtained by classic PCR and by in situ PCR of frozen sections.
Conclusion:
In situ PCR on smears could be used in targeted-screening for young and post-menopausal women, as well as in the development of large scale studies to establish the connection among the presence of HPV, p53 polymorphisms and the risk of cervical cancer.
Abbreviations:
PCR, polymerase chain reaction; OsO4, osmium tetroxide; HPV, human papilloma virus; PBS, phosphate-buffered saline; SDS, sodium dodecyl sulphate.
