HPV Viral Activity by mRNA-HPV Molecular Analysis to Screen the Transforming Infections in Precancer Cervical Lesions

Andrea      Tinelli; Giuseppe      Leo; Maurizio      Pisano; Fabio      Storelli; Sara      Leo; Daniele      Vergara; Antonio      Malvasi

Abstract

Introduction: a current problem with the Human papillomavirus (HPV) genital infection is to detect HPV presence and activity in high risk women. Material and methods: 190 women at risk for HPV-infection underwent a Pap Test as well as a cervico-vaginal mucus sample analysis. The genome amplification of ORF L1 was by REAL-Time PCR by direct sequencing in capillary elettrophoresis of amplified product in Real Time (by ABI PRISM® 310 Genetic Analyzer, Applied Biosystem, USA), followed by HPV genotyping using oligonucleotide probe hybridization. Degenerate primers My09/11, with 450 bp product amplified were utilized in Real Time and in Direct Sequencing. Furthermore, samples were evaluated by mRNA-HPV test to detect the presence of E6 and E7 transcripts. The results were compared with cytology. Results: a total of 62 women were positive for HPV infection (32.6%) and 19 of these had one or more high-risk HPVs (30.6%); the concordance between the two assays was 78.9%, with 21.1% of totally or partially discordant results. Cytological results showed mRNA presence in 4 low grade and 2 high grade squamous intraepithelial lesions. Conclusion: the results suggest the potential of E6/E7 detection to target the presence of a transforming HPV infection.

Keywords: HPV, Human papillomavirus, high-risk HPV, cervical cancer, DNA, mRNA, NASBA, REAL-Time PCR squamous intraepithelial lesion, SIL, cervical intraepithelial neoplasia