HPV E6/E7 mRNA vs. HPV DNA in co-testing with ThinPrep cytology for cervical cancer screening: a prospective observational diagnostic accuracy study of methylation biomarkers.

OBJECTIVE: Human papillomavirus (HPV) gene transcription activity and methylation could be promising tools for cervical cancer (CC) screening. This study compared HPV E6/E7 messenger RNA (mRNA) + ThinPrep cytology test (TCT) vs. HPV genotyping + TCT in screening CC and explored the relationship between gene methylation and cervical lesions.

METHODS: Women who visited gynecology outpatient clinics between January 2023 and November 2024 were enrolled and assigned to 2 screening strategies based on their wishes: HPV DNA + TCT or HPV E6/E7 mRNA + TCT. Methylation testing of FAM19A4, ZNF582, EPHA7, SOX17, PAX1, and JAM3 genes was performed on the biopsy specimens from 100 patients.

RESULTS: Ultimately, 1,399 and 991 women underwent HPV DNA + TCT and HPV E6/E7 mRNA + TCT screening, respectively. The sensitivity, specificity, and accuracy in diagnosing high-grade squamous intraepithelial lesion (HSIL)+ were 72.09%, 83.05%, and 81.7% for HPV DNA + TCT, and 77.97%, 86.37%, and 85.37% for HPV E6/E7 mRNA + TCT, respectively. The areas under the receiver operating characteristic curves were 0.777 for HPV DNA + TCT and 0.822 for HPV E6/E7 mRNA + TCT. No HSIL+ lesions were observed in 65 patients with HPV 16/18 (+) but HPV E6/E7 mRNA (-). In cases with HPV 16/18 (+) and HPV E6/E7 mRNA (+), the immediate risks for cervical intraepithelial neoplasia (CIN) 2+ and CIN3+ were 64.06% and 50%, respectively. Methylation levels of FAM19A4, ZNF582, EPHA7, SOX17, PAX1, and JAM3 genes increased with lesion severity.

CONCLUSION: The sensitivity and specificity of HPV E6/E7 mRNA + TCT appeared higher than HPV DNA + TCT. Simultaneous positivity for HPV 16/18 and HPV E6/E7 mRNA suggests a high immediate risk of CIN2+ and CIN3+. These results may improve CC screening.