Detection of oncogenic HPV and identification of 72Arg polymorphic p53 by in situ PCR for clinical routine purposes.
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.


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.


CONCLUSIONS
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.


BACKGROUND
PCR, polymerase chain reaction; OsO4, osmium tetroxide; HPV, human papilloma virus; PBS, phosphate-buffered saline; SDS, sodium dodecyl sulphate.
Realacci
Massimo
M
Department of Experimental Medicine and Pathology, University La Sapienza, Rome, Italy.
Perrone
Giulietta Anna
GA
Sale
Patrizio
P
Tafani
Marco
M
Di Vito
Maura
M
Perrone-Donnorso
Raffaele
R
Zotti
Giancarlo
G
Russo
Matteo A
MA
D005260:Female; D016158:Genes, p53; D052162:Human papillomavirus 16 / Q000145:classification / Q000235:genetics*; D052161:Human papillomavirus 18 / Q000145:classification / Q000235:genetics*; D006801:Humans; D030361:Papillomavirus Infections / Q000235:genetics* / Q000473:pathology / Q000821:virology*; D011110:Polymorphism, Genetic; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D016159:Tumor Suppressor Protein p53 / Q000235:genetics*; D014626:Vaginal Smears
C495901:TP53 protein, human; D016159:Tumor Suppressor Protein p53