Detection of Oncogenic HPV and Identification of 72^Arg Polymorphic p53 by In Situ PCR for Clinical Routine Purposes

Abstract

Background: Ano-genital carcinoma is a poly-factorial 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.