Int J Gynecol Obstet · 2025;171(1):405–414 · METHY2/3 · China

Cytologic DNA Methylation for Minimally Abnormal Screening Results

Cytologic DNA methylation for managing minimally abnormal cervical cancer screening results
1,857 minimally abnormal women (hrHPV+/NILM, ASC-US, LSIL) · CIN3+ Se 74.9%, Sp 89.1% · CIN3+ risk: 40.5% if CISCER(+) vs 2.7% if (−)
Shang X, Kong L, You Y, Wu H, Liou Y, Jin X, Liu P, Lang J, Li L
Peking Union Medical College Hospital · Int J Gynecol Obstet 2025;171(1):405–414 · doi:10.1002/ijgo.70167
1,857
minimally abnormal results
40.5%
CIN3+ risk if CISCER(+)
2.7%
CIN3+ risk if CISCER(−)
0.820
AUC · CIN3+

1Background & Objective

  • Problem: hrHPV+ with normal cytology, ASC-US or LSIL — complex algorithms, excessive colposcopy referral.
  • Objective: test CISCER (PAX1/JAM3 methylation) for managing minimally abnormal results.

2Study Design & Cohort

METHY2
NCT03960879
METHY3
NCT04646954
2019–22
enrollment
  • Design: cross-sectional, colposcopy-referred, PUMCH; histology as gold standard.
  • Inclusion: women with minimally abnormal screening results & histologic outcome.
  • Minimally abnormal: hrHPV+ / NILM · ASC-US · LSIL.
  • Outcome: CIN3+ detection; absolute risk & referrals within one screening round.
  • Analysis: ROC/AUC; sensitivity & specificity; immediate risk stratification.
13.72
ΔCt PAX1 · normal
5.24
ΔCt PAX1 · CIN3
3.74
ΔCt PAX1 · cancer
15.08
ΔCt JAM3 · normal
8.24
ΔCt JAM3 · CIN3
5.85
ΔCt JAM3 · cancer
167
CIN3+ cases
9.0%
baseline CIN3+ risk
11.1
referrals / CIN3+ (no triage)
  • Inclusion: ≥18 y, intact cervix, NILM/ASC-US/LSIL cytology, full consent.
  • Exclusion: pregnancy · HIV+ · transplant on antirejection.
  • Samples: cervical cytology → hrHPV DNA · cytology · CISCER methylation.
  • Referral: per ASCCP/China guidelines; methylation not an indication.
  • Statistics: Mann–Whitney U · χ² · ROC; risk with 95% CI.
  • Readout: CISCER(+): ΔCt PAX1 ≤6.6 or JAM3 ≤10.0.
Lower ΔCt = higher methylation. Methylation positive was NOT a colposcopy indication — tested independently.

3CISCER Performance — CIN3+

ROC curves of CISCER vs single genes
Fig. ROC — CISCER AUC 0.820 vs PAX1 0.778 & JAM3 0.777.
CIN3+ risk before and after CISCER triage
Fig. Pre-/post-test CIN3+ risk — referral reduction per one CIN3+ found (11.1 → 2.5).
74.9%
Sensitivity
89.1%
Specificity
97.3%
NPV
0.820
AUC CIN3+
  • Superior to single genes: CISCER AUC 0.820 > PAX1 0.778 / JAM3 0.777.
  • Age >50 y: Se 79.5% / Sp 89.3% — high accuracy in older women.

4Performance by Cytology & Age

NILM
AUC 0.867
ASC-US
AUC 0.812
LSIL
AUC 0.787
Age >50 y
higher AUC & Se

5Risk Stratification & Referral Reduction

13.7→3.7
ΔCt PAX1 · normal→cancer
15.1→5.9
ΔCt JAM3 · normal→cancer
40.5%
CIN3+ risk · CISCER(+)
2.7%
CIN3+ risk · CISCER(−)
11.1→2.5
referrals per CIN3+ found
  • Risk gradient 15×: 40.5% vs 2.7% — clear positive/negative separation.
  • Actionable: CISCER(+) → colposcopy; CISCER(−) → defer & re-screen.
Clinical Significance