1Background & Objective
- Problem: hrHPV screening is sensitive but low-specificity; cytology triage is subjective & poorly reproducible.
- Objective: multicenter prospective validation of PAX1m/JAM3m vs TCT & hrHPV for CIN2+/CIN3+.
2Study Design & Cohort
3
centers
2022.5–10
window
39y
median age
- Design: prospective multicenter; opportunistic screening; histology reference.
- Assays: TCT + Cobas hrHPV + PAX1m/JAM3m (PCR-fluorescence).
- Readout: ΔCt PAX1 ≤ 6.6 or ΔCt JAM3 ≤ 10.0.
45.7%
normal / cervicitis
23.1%
CIN1
14.2%
CIN2
11.8%
CIN3
5.2%
cancer
- Inclusion: ≥18 y, intact cervix; no severe immunodeficiency.
- Exclusion: known genital malignancy or active other cancers.
- Flow: guideline-based colposcopy & biopsy; blinded methylation.
- Compare: single & combined strategies for CIN2+/CIN3+.
0.872
methylation AUC
0.580
TCT AUC
0.503
hrHPV AUC
Ethics KS2021211 · ClinicalTrials.gov NCT05290428.
3Dual-Endpoint Performance
87.6%
Se · CIN3+
86.8%
Sp · CIN3+
74.1%
Se · CIN2+
95.9%
Sp · CIN2+
- Accuracy: excellent for both CIN2+ and CIN3+ — superior to traditional screening.
- Specificity edge: 95.9% (CIN2+) — far fewer false referrals than cytology/HPV.
4AUC — CIN3+ (the decisive comparison)
All P<0.05 — methylation AUC significantly higher.
5Triage Implications
1,184
histology-confirmed
62
cancers
3
centers
- hrHPV+ triage: methylation as reflex test — maintains sensitivity, raises specificity, cuts colposcopy load.
- Objective: PCR-fluorescence — reproducible, operator-independent.
- Multicenter evidence: validated across 3 Chinese centers — generalizable to opportunistic screening.
- Cancer safety: methylation positive in all cancers in this cohort.
- Dual endpoint: strong for both CIN2+ and CIN3+ — screening-ready.
- Implementation: same cervical sample as TCT/HPV — easy integration.
- Cost-benefit: fewer unnecessary colposcopies offset assay cost.
Methylation complements cytology & HPV — an objective upgrade to the screening pathway.