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Genotype-Specific HPV E6/E7 mRNA Triage for Risk Stratification in HPV DNA-Positive Women with ASC-US/LSIL: A
Sveinung Wergeland Sørbye1, Bente Marie Falang2, Mona Antonsen1
1Department of Clinical Pathology, University Hospital of North Norway, 9006 Tromsø, Norway.
Pathogens (Basel, Switzerland)
|July 28, 2026
Summary
Genotype-specific HPV E6/E7 mRNA testing effectively stratifies risk in HPV DNA-positive women with ASC-US/LSIL cytology. This approach improves cervical cancer screening accuracy and can reduce unnecessary colposcopy referrals.
Area of Science:
- Gynecology
- Oncology
- Virology
Background:
- Human papillomavirus (HPV) DNA-positive women with atypical squamous cells of undetermined significance (ASC-US) or low-grade squamous intraepithelial lesion (LSIL) cytology form a heterogeneous group requiring effective triage.
- Current triage methods may not optimally identify individuals at high risk for cervical precancer or cancer.
Purpose of the Study:
- To evaluate the clinical utility of genotype-specific HPV E6/E7 mRNA testing for risk stratification in HPV DNA-positive women with ASC-US/LSIL cytology.
- To compare the performance of mRNA testing with HPV DNA testing in predicting cervical intraepithelial neoplasia (CIN2+ and CIN3+).
Main Methods:
- A retrospective quality-assurance study involving 1006 HPV DNA-positive women with ASC-US/LSIL cytology screened between 2019 and 2024.
- Genotype-specific HPV E6/E7 mRNA testing using the PreTect HPV-Proofer assay, detecting mRNA from HPV types 16, 18, 31, 33, 45, 52, and 58.
- Linkage to regional pathology records to determine CIN2+ and CIN3+ diagnoses.
Main Results:
- ASC-US/LSIL cytology represented 42.5% of HPV DNA-positive women in the Tromsø cohort.
- In the mRNA-tested cohort, CIN2+ was significantly higher in mRNA-positive women (26.1%) compared to mRNA-negative women (7.9%), with a risk ratio of 3.31.
- The assay demonstrated a sensitivity of 69.5% and specificity of 64.4% for CIN2+.
- Simulated mRNA-guided referral could reduce immediate colposcopy referrals by 59.2%.
Conclusions:
- Genotype-specific HPV E6/E7 mRNA testing provides clinically relevant risk stratification for HPV DNA-positive women with ASC-US/LSIL cytology.
- mRNA testing shows promise in identifying women with a higher risk of CIN2+, thereby optimizing colposcopy referrals.
- While mRNA testing aids risk assessment, structured surveillance remains essential for mRNA-negative women.
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