THE CUTOFF VALUE OF THE E6/E7 mRNA EXPRESSION RATE BY FLOW CYTOMETRY FOR THE DETECTION OF CERVICAL CYTOLOGICAL ABNORMALITIES
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Abstract
Background: Persistent infection with high-risk Human Papillomavirus (HR-HPV) types is the main risk factor for cervical cancer. The expression of the viral oncogenes E6 and E7 reflects the biological activity of HPV and is associated with the progression of cervical lesions. Therefore, assessing E6/E7 mRNA expression levels may serve as a potential biomarker for detecting cervical cytological abnormalities.
Objective: To determine the optimal cutoff value of E6/E7 mRNA expression rate for detecting cervical cytological abnormalities.
Methods: A cross-sectional descriptive study was conducted on 527 women undergoing cervical cancer screening at Dai Phuoc General Clinic from January 2023 to May 2025. All liquid-based cervical cytology samples were simultaneously tested using cytology, HR-HPV DNA detection by multiplex real-time PCR, and quantification of E6/E7 mRNA expression using flow cytometry. The optimal cutoff value was determined using ROC curve analysis, with cytology as the reference standard. Agreement between methods was evaluated using Gwet’s AC coefficient.
Results: The optimal cutoff for E6/E7 mRNA expression was 0,15%, with sensitivity 58,82%, specificity 85,29%, and accuracy 84,44%; AUC = 0,69 (95% CI: 0,52–0,86; p = 0,026). Detection of E6/E7 mRNA showed good agreement with cytology (Gwet’s AC = 0,80; 95% CI: 0,75–0,84; p < 0,001).
Conclusion: An E6/E7 mRNA expression cutoff of 0,15% demonstrated good specificity and strong agreement with cytology for detecting cervical cytological abnormalities, indicating its potential utility in cervical cancer screening and risk stratification.
Keywords
HR-HPV, mRNA E6/E7, Flow cytometry.
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References
2. Bedell SL, Goldstein LS, Goldstein AR, Goldstein AT. Cervical Cancer Screening: Past, Present, and Future. Sexual Medicine Reviews. 2020;8(1):28-37.
3. Baba SK, Alblooshi SSE, Yaqoob R, Behl S, Al Saleem M, Rakha EA, et al. Human papilloma virus (HPV) mediated cancers: an insightful update. Journal of Translational Medicine. 2025;23(1):483.
4. Derbie A, Mekonnen D, Woldeamanuel Y, Van Ostade X, Abebe T. HPV E6/E7 mRNA test for the detection of high grade cervical intraepithelial neoplasia (CIN2+): a systematic review. Infectious agents and cancer. 2020;15:9.
5. Pan C, Zhou J, Lyu J, Ren X. Development and validation of a multiplex reverse transcript real-time PCR for E6/E7 mRNA detection of high-risk human papillomavirus. Journal of medical microbiology. 2018;67(10):1509-14.
6. Wongpakaran N, Wongpakaran T, Wedding D, Gwet KL. A comparison of Cohen's Kappa and Gwet's AC1 when calculating inter-rater reliability coefficients: a study conducted with personality disorder samples. BMC medical research methodology. 2013;13:61.
7. Zhu H, Shen Z, Luo H, Zhang W, Zhu X. Chlamydia Trachomatis Infection-Associated Risk of Cervical Cancer: A Meta-Analysis. Medicine. 2016;95(13):e3077.
8. Chen L, Wang H. eIF4E is a critical regulator of human papillomavirus (HPV)-immortalized cervical epithelial (H8) cell growth induced by nicotine. Toxicology. 2019;419:1-10.
9. Kottaridi C, Tsiodras S, Spathis A, Chranioti A, Pappas A, Kassanos D, et al. Clinical performance of human papillomavirus E6, E7 mRNA flow cytometric assay compared to human papillomavirus DNA typing. Analytical and quantitative cytology and histology. 2011;33(6):305-10.
10. Pierry D, Weiss G, Lack B, Chen V, Fusco J. Intracellular human papillomavirus E6, E7 mRNA quantification predicts CIN 2+ in cervical biopsies better than Papanicolaou screening for women regardless of age. Archives of pathology & laboratory medicine. 2012;136(8):956-60.
11. Gupta SM, Warke H, Chaudhari H, Mavani P, Katke RD, Kerkar SC, et al. Human Papillomavirus E6/E7 oncogene transcripts as biomarkers for the early detection of cervical cancer. Journal of medical virology. 2022;94(7):3368-75.