PAX1m/JAM3m Methylation for Triage of ASC-US — Chen et al. 2024, BMC Cancer

1Background & Objective

ASC-US burden: >50% of cytological abnormalities in China; <30% harbor high-grade lesions — over/under-diagnosis risk.
Triage gap: cytology low sensitivity; HPV referral rate 91.1% with only 10.8% specificity for CIN2+.
Objective: evaluate CISCER® PAX1m/JAM3m for triaging women with ASC-US.
Why methylation: objective, reproducible, HPV-independent marker of malignant transformation.

2Study Design & Cohort

322

ASC-US women

203

with histology

CIN2+ (20 CIN2 · 16 CIN3 · 1 AdCa)

HPV-DNA+ cytology ASC-US+ PAX1m/JAM3m

Setting: Zhejiang Provincial People's Hospital, 2022.3–2024.1; ethics No. 2021SJ020.

3Methylation Signal by Grade

Dose–response: ΔCt falls sharply from CIN1 to CIN2 (P<0.001); stays low in CIN3; adenocarcinoma ΔCt_PAX1=1.78, ΔCt_JAM3=3.4.
Low-grade specificity: in no-CIN/CIN1, PAX1 or JAM3 positive only 2.4% (4/166).
Positivity rises: CIN2 50% both genes; CIN3 50% both genes (Fig. 2C).

2.4%

positive · no-CIN/CIN1

80%

positive · CIN2

87.5%

positive · CIN3

4Results — ROC, Forest & Referral

ROC of different triage methods for ASC-US patients with CIN2+

Fig. 3 ROC of triage methods for CIN2+ in ASC-US.

Forest plots of relative specificity and sensitivity

Fig. 4 Forest: relative specificity of methylation 8.84× HPV (6.35–15.18) at comparable sensitivity.

HPV triage

91.1%

CISCER triage

18.7%

Referral: HPV 91.1% → CISCER 18.7%; in hr-HPV+ women −79.5% (147/185).
Risk stratification: CIN3+ risk CISCER(+) 39.5% vs CISCER(−) 1.2%.
Best in non-16/18: CISCER Se 90.5% (vs 75.0% in HPV16/18+).

39.5%

CIN3+ risk · CISCER(+)

1.2%

CIN3+ risk · CISCER(−)

Subgroup	Se %	Sp %
Non-16/18 hrHPV+	90.5	95.6
HPV16/18+	75.0	94.1

5Performance — CIN2+ (n=37)

Test	Se %	Sp %	PPV %	NPV %	OR
CISCER	83.8	95.8	81.6	96.4	121.1
PAX1m	70.3	97.6	86.7	93.6	95.7
JAM3m	64.8	96.4	80.0	92.5	51.3
HPV	100	10.8	20	100	—

CISCER = PAX1m or JAM3m positive (ΔCt PAX1 ≤ 6.6 / ΔCt JAM3 ≤ 10.0).

6Referral & Safety

91.1% → 18.7%

Referral rate cut 79.5% in hr-HPV+

without missing CIN2+ (HPV16/18 & non-16/18 groups)

39.5%

CIN3+ risk if CISCER(+)

vs 1.2% if CISCER(−) — negative results are safe to follow up

18.7%

Whole-cohort referral rate

vs 91.1% for HPV — 4.9× fewer colposcopies

7Who Benefits Most

40y

median age

HPV16/18+

206

non-16/18 hrHPV+

Non-16/18 hrHPV+: CISCER Se 90.5%, Sp 95.6% — best triage effect.
HPV16/18+: CISCER Se 75.0%, Sp 94.1% — still reduces referral burden.
HPV-negative ASC-US: all 18 were CIN1− and CISCER− — safe to follow up.