PAX1m/JAM3m Triage in Non-16/18 hrHPV+ Women — Chen et al. 2024, Clinical Epigenetics

1Background & Objective

Burden: non-16/18 hrHPV types drive much HSIL+/cancer in Asia; triage still relies on cytology.
Cytology limits: subjective, lab-dependent, low specificity (26.7%) → colposcopy overreferral.
Objective: validate CISCER® PAX1m/JAM3m for CIN3+ triage in non-16/18 hrHPV+ women.

2Study Design & Cohort

30,084

screened · 6 hospitals

→

4,735

non-16/18 hrHPV+

→

1,851

with histology

prospective cohorts

hospitals

42y

median age

HPV genotyping+ LBC cytology+ PAX1m/JAM3m

METHY3 · 2020.11–2021.12

14 mo

METHY4 · 2022.05–2022.10

6 mo

Cohorts: METHY3 + METHY4, opportunistic screening; lab blinded.
Endpoint: CIN3+ detection; comparator LBC ≥ASC-US; colposcopy/surgery histology.
Statistics: ROC/AUC, McNemar; SPSS 26 / R 4.1.2.
Inclusion: ≥18 y, intact cervix, no severe immunodeficiency.
Assay: bisulfite conversion + qPCR (GAPDH control, ABI 7500).
Outcome: 151 CIN3+ (131 CIN3 + 20 cancers) among 1,851.

82.3%

Normal/CIN1

9.6%

CIN2

7.1%

CIN3

1.1%

Cancer (n=20)

3Key Results — Signal & Performance

PAX1/JAM3 ΔCt distributions by pathology and cytology

Fig. 2 PAX1/JAM3 ΔCt by histology & LBC grade — rises with severity (P<0.05).

Fig. 3 Se vs Sp by triage (data: Table 2) — methylation: high both; cytology: high Se, very low Sp.

Fig. 4 OR for CIN3+ (Table 3) — P/J(+) 42.6 vs LBC(+) 3.3.

Dose–response: ΔCt falls as lesions progress — all 20 cancers were PAX1m/JAM3m(+); positive = ΔCt PAX1 ≤ 6.6 or ΔCt JAM3 ≤ 10.0.

4CIN3+ Detection & Referral Burden

Method	Sens. %	Spec. %	AUC	NPV %
PAX1m/JAM3m (+)	84.8	88.5	0.866	98.5
LBC ≥ ASC-US	90.1	26.7	0.584	96.8
LBC or P/J (+)	98.0	24.8	0.614	99.3

OR for CIN3+: P/J(+) 42.6 (27.1–69.6) vs LBC(+) 3.3 (2.0–5.9), P<0.001.

Triage	Referred	% of 1,851
PAX1m/JAM3m (+)	323	17.45%
LBC ≥ ASC-US	1,382	74.66%
LBC or P/J (+)	1,427	77.09%

Methylation alone keeps the lowest referral burden.

LBC ≥ ASC-US

74.7%

PAX1m/JAM3m

17.5%

Referral rate — P/J(+) triage cuts referrals by 57.2%.

5Cancer Safety & Risk-Based Management

20 / 20

cancers caught by P/J(+)

100%

16 / 20

caught by LBC ≥ASC-US

80% — 4 missed

42.6

OR for CIN3+ if P/J(+)

98.5%

NPV if P/J(−)

Missed by cytology: 3 SCC + 1 ADC — all four were P/J(+).
P/J(+) → direct colposcopy (high CIN3+ odds); P/J(−) → safe follow-up (NPV 98.5%).
Standalone: adding cytology gives no triage advantage — methylation alone is the better strategy.