HPV viral activity by mRNA-HPV molecular analysis to screen the transforming infections in precancer cervical lesions.
BACKGROUND
a current problem with the Human papillomavirus (HPV) genital infection is to detect HPV presence and activity in high risk women.


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((R)) 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.


CONCLUSIONS
the results suggest the potential of E6/E7 detection to target the presence of a transforming HPV infection.
Tinelli
Andrea
A
Department of Obstetrics and Gynaecology, Vito Fazzi Hospital, Lecce, Italy. andreatinelli@gmail.com
Leo
Giuseppe
G
Pisanò
Maurizio
M
Storelli
Fabio
F
Leo
Sara
S
Vergara
Daniele
D
Malvasi
Antonio
A
D000328:Adult; D015331:Cohort Studies; D015344:DNA Probes, HPV; D005260:Female; D006801:Humans; D009693:Nucleic Acid Hybridization; D027383:Papillomaviridae / Q000235:genetics / Q000302:isolation & purification*; D030361:Papillomavirus Infections / Q000453:epidemiology / Q000473:pathology / Q000821:virology*; D011230:Precancerous Conditions / Q000453:epidemiology / Q000473:pathology / Q000821:virology*; D012333:RNA, Messenger / Q000235:genetics*; D012367:RNA, Viral / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D002583:Uterine Cervical Neoplasms / Q000453:epidemiology / Q000473:pathology / Q000821:virology*; D014626:Vaginal Smears; D002578:Uterine Cervical Dysplasia / Q000453:epidemiology / Q000473:pathology / Q000821:virology*
D015344:DNA Probes, HPV; D012333:RNA, Messenger; D012367:RNA, Viral