Diagnostic performance of HPV E6/E7 mRNA testing towards HPV-DNA testing and p16/Ki67 immunostaining as a biomarker of high-risk HPV recurrence in Greek women surgically treated for their cervical lesions.

AIM: To evaluate the diagnostic performance of E6/E7 HPV-mRNA overexpression towards HPV-DNA testing and p16/Ki67 immunocytochemistry in a post-op population to verify if this biomarker can be effectively used as indicator of successful cervical intraepithelial neoplasia (CIN) treatment. METHODS: Our study retrospectively analyzed 197 patients of our Colposcopy Clinic between January 2013 and September 2020 coming with an abnormal Pap smear suggestive for colposcopy, and after a series of follow-ups including liquid-based cytology (LBC) and punch-biopsy sampling, there were surgically treated. LBC was used for cytology and molecular analysis of the three HPV-related biomarkers. RESULTS: Six months after treatment, 93% of the HPV-mRNA-positive women became negative while this applied to only 80.2% of the HPV-DNA-positive women. HPV persistence was 6.9% at 6-12 months after treatment. The comparison among cytology, colposcopy, HPV-DNA test, and HPV-mRNA test after treatment revealed that the last one is the only with a strong correlation with actual severity (histology during treatment) (ρ = 0.345, p = 0.006) implying that clinical cases with more severe CIN may have higher chances of unsuccessful treatment. HPV-mRNA test had higher sensitivity (100%), specificity (96.88%), and positive predictive value (45.45%) for CIN2+ recurrent lesions when compared with HPV-DNA testing (80%, 82.81%, 10.81% respectively) and p16/Ki67 immunocytochemistry (80%, 95.83%, 33.33% respectively) while their negative predictive values were similar. CONCLUSIONS: E6/E7 mRNA detection has higher diagnostic values for the prediction of treatment failure compared with HPV-DNA testing and p16/Ki67 immunocytochemistry, and as an outcome could be used as predictive indicator of CIN-treatment status.

A cohort retrospective study of high-risk HPV recurrence in Greek women after cervical lesion treatment through detection of viral E6/E7 mRNA expression.

PURPOSE: Our aim was to detect and evaluate potential alterations in the postoperative status of E6/E7 HPV mRNA in women treated for cervical intraepithelial lesions (CIN) and if so, to evaluate its potential use as a prognostic tool to identify patients with increased risk of treatment failure or recurrent disease. METHODS: Our study retrospectively analyzed 101 women with an abnormal Pap smear, or in some cases with histological reports or molecular analysis requiring colposcopic evaluation. Thin-prep cytological samples were collected before colposcopy and histology (when necessary). After treatment, all women were scheduled for colposcopy in six months. The cytological material was analyzed with CLART-2 HPV-DNA test and HPV-PROOFER E6/E7 mRNA test. RESULTS: Concerning demographics, no significant correlations were found for smoking, condom use or vaccination status. It seems that the only statistically significant correlation with actual severity came from the mRNA-test after treatment. This shows that clinical cases with more severe CIN may have higher chances of unsuccessful treatment. At the first post-op visit, 83.5% of HPV mRNA-positive women had a negative HPV mRNA-test while only 60.4% of HPV DNA-positive women became negative. There were 12 HPV-mRNA positive patients both before and after treatment, 3 of whom had a negative HPV DNA test, meaning that, if based only on HPV-DNA results, they would have been managed wrongly as successfully treated patients. Our study shows that E6/E7 mRNA detection has particularly high specificity and positive likelihood ratio for the prediction of treatment failure in comparison with HPV DNA-testing. CONCLUSIONS: E6/E7 mRNA overexpression seems to be a promising candidate as an indicator-biomarker to determine the success of treatment.