Toward the simultaneous detection of multiple diseases with a highly cost-effective cell-free DNA methylome test

W Weihua Zeng (Department of Pathology and Laboratory Medicine, David Geffen School of Medicine, University of California at Los Angeles) C Chun-Chi Liu (EarlyDiagnostics Inc.) S Shuo Li Y Yonggang Zhou (Department of Pathology and Laboratory Medicine, David Geffen School of Medicine, University of California at Los Angeles) M Mary L. Stackpole (EarlyDiagnostics Inc.) Y Ying Xiao (CIBM Center for Biomedical Imaging) R Ran Hu (State Key Laboratory of Water Resources Engineering and Management, Wuhan University) C Caitlin Tang (Department of Pathology and Laboratory Medicine, David Geffen School of Medicine, University of California at Los Angeles) Q Qiao Liu W Wanwen Zeng (Department of Statistics, Stanford University) A Angela Yeh (Department of Medicine, David Geffen School of Medicine, University of California at Los Angeles) A Andrew Melehy (Department of Surgery, David Geffen School of Medicine, University of California at Los Angeles) B Benjamin Tran (Department of Surgery, David Geffen School of Medicine, University of California at Los Angeles) Z Zorawar Noor (Department of Medicine, David Geffen School of Medicine, University of California at Los Angeles) M Megumi Yokomizo (Department of Radiological Sciences, David Geffen School of Medicine, University of California at Los Angeles) D Dominic Amara (Department of Surgery, David Geffen School of Medicine, University of California at Los Angeles) S Shreya Gumate (Department of Surgery, David Geffen School of Medicine, University of California at Los Angeles) P Preeti Ahuja (Department of Radiological Sciences, David Geffen School of Medicine, University of California at Los Angeles) D David Yuanze Li (Thousand Oaks High School) J Junting Zhao (Sage Hill School) I Inga Rose (Reference Medicine) C Cattlena Walker (Crown Bioscience) S Sadaf Malik (Crown Bioscience) Y Yazhen Zhu (Department of Pathology and Laboratory Medicine, David Geffen School of Medicine, University of California at Los Angeles) H Hsian-Rong Tseng (Jonsson Comprehensive Cancer Center, University of California at Los Angeles) E Edward B. Garon S Samuel W. French (Department of Pathology and Laboratory Medicine, David Geffen School of Medicine, University of California at Los Angeles) C Clara E. Magyar (Department of Pathology and Laboratory Medicine, David Geffen School of Medicine, University of California at Los Angeles) S Sarah M. Dry (Department of Pathology and Laboratory Medicine, David Geffen School of Medicine, University of California at Los Angeles) C Clara M. Lajonchere (Jonsson Comprehensive Cancer Center, University of California at Los Angeles) D Daniel Geschwind G Gina Choi (Department of Medicine, David Geffen School of Medicine, University of California at Los Angeles) S Sammy Saab (Department of Medicine, David Geffen School of Medicine, University of California at Los Angeles) A Akshay Shetty (Department of Medicine, David Geffen School of Medicine, University of California at Los Angeles) C Carrie R. Wong (Vatche and Tamar Manoukian Division of Digestive Diseases, Department of Medicine, University of California) K Kevin G. King (Department of Radiological Sciences, David Geffen School of Medicine, University of California at Los Angeles) D David S. Lu (Department of Radiological Sciences, David Geffen School of Medicine, University of California at Los Angeles) S Steven S. Raman (Department of Radiological Sciences, David Geffen School of Medicine, University of California at Los Angeles) X Xiyan Xiang K Kirti Shetty (Division of Gastroenterology and Hepatology, School of Medicine, University of Maryland) L Lopa Mishra S Sanaz Memarzadeh (Jonsson Comprehensive Cancer Center, University of California at Los Angeles) Y Yan Liu F Frank Alber W William Hsu (Department of Radiological Sciences, David Geffen School of Medicine, University of California at Los Angeles) K Kostyantyn Krysan (Department of Medicine, David Geffen School of Medicine, University of California at Los Angeles) S Steven M. Dubinett D Denise R. Aberle (Department of Radiological Sciences, David Geffen School of Medicine, University of California at Los Angeles) V Vatche Agopian (Department of Surgery, David Geffen School of Medicine, University of California at Los Angeles) S Steven-Huy B. Han (Department of Medicine, David Geffen School of Medicine, University of California at Los Angeles) W Wing Hung Wong (Department of Statistics, Stanford University) X Xiaohui Ni (EarlyDiagnostics Inc.) W Wenyuan Li (Department of Biochemistry, The University of Texas Southwestern Medical Center, 5323 Harry Hines Blvd., Dallas, Texas 75390, United States) X Xianghong Jasmine Zhou (Department of Pathology and Laboratory Medicine, David Geffen School of Medicine, University of California at Los Angeles)

Abstract

Plasma cell-free DNA (cfDNA), originating from multiple organs, holds significant potential for noninvasive diagnostics and prognostics. Current cfDNA methylation assays primarily focus on single clinical indications by targeting specific genomic loci. In contrast, comprehensive profiling of cfDNA methylome can enable simultaneous detection of multiple diseases by capturing organ-specific methylation signatures, thereby offering a holistic view of health, when disease etiology is unclear or when conventional biochemical diagnostics are unavailable. However, deep sequencing required for sensitive detection of methylation abnormalities remains prohibitively expensive, limiting widespread clinical use. To overcome this barrier, we developed MethylScan , a highly cost-effective approach for cfDNA methylome sequencing. We demonstrated its broad clinical utility in a cohort of 1,061 individuals across diverse applications, including multicancer detection in general population, liver cancer surveillance in high-risk individuals, liver disease classification, identification of organ abnormalities, and race prediction from cfDNA. In multicancer detection (liver, lung, ovarian, and stomach cancers), MethylScan achieved an area under the receiver operating characteristic curve (AUROC) of 0.938 (95% CI: 0.920 to 0.954), with a sensitivity of 63.3% (95% CI: 58.9 to 67.9%) at 98.0% specificity for all cancer stages. For early-stage cancers, the AUROC was 0.916 (95% CI: 0.890 to 0.940), with 55.3% sensitivity (95% CI: 49.1 to 62.1%) at the same specificity. In liver cancer surveillance, MethylScan achieved an AUROC of 0.927 (95% CI: 0.889 to 0.959), with 79.6% sensitivity (95% CI: 70.6 to 87.8%) at 90.4% specificity. The assay also demonstrated strong performance in additional diagnostic tasks, supporting its potential as a versatile platform for comprehensive cfDNA-based health monitoring.

Article Details

Volume / Issue Vol. 123, Issue 15
Published April 14, 2026
ISSN 0027-8424
Publisher National Academy of Sciences

Authors (54)

W

Weihua Zeng

Department of Pathology and Laboratory Medicine, David Geffen School of Medicine, University of California at Los Angeles

C

Chun-Chi Liu

EarlyDiagnostics Inc.

S

Shuo Li

Y

Yonggang Zhou

Department of Pathology and Laboratory Medicine, David Geffen School of Medicine, University of California at Los Angeles

M

Mary L. Stackpole

EarlyDiagnostics Inc.

Y

Ying Xiao

CIBM Center for Biomedical Imaging

R

Ran Hu

State Key Laboratory of Water Resources Engineering and Management, Wuhan University

C

Caitlin Tang

Department of Pathology and Laboratory Medicine, David Geffen School of Medicine, University of California at Los Angeles

Q

Qiao Liu

W

Wanwen Zeng

Department of Statistics, Stanford University

A

Angela Yeh

Department of Medicine, David Geffen School of Medicine, University of California at Los Angeles

A

Andrew Melehy

Department of Surgery, David Geffen School of Medicine, University of California at Los Angeles

B

Benjamin Tran

Department of Surgery, David Geffen School of Medicine, University of California at Los Angeles

Z

Zorawar Noor

Department of Medicine, David Geffen School of Medicine, University of California at Los Angeles

M

Megumi Yokomizo

Department of Radiological Sciences, David Geffen School of Medicine, University of California at Los Angeles

D

Dominic Amara

Department of Surgery, David Geffen School of Medicine, University of California at Los Angeles

S

Shreya Gumate

Department of Surgery, David Geffen School of Medicine, University of California at Los Angeles

P

Preeti Ahuja

Department of Radiological Sciences, David Geffen School of Medicine, University of California at Los Angeles

D

David Yuanze Li

Thousand Oaks High School

J

Junting Zhao

Sage Hill School

I

Inga Rose

Reference Medicine

C

Cattlena Walker

Crown Bioscience

S

Sadaf Malik

Crown Bioscience

Y

Yazhen Zhu

Department of Pathology and Laboratory Medicine, David Geffen School of Medicine, University of California at Los Angeles

H

Hsian-Rong Tseng

Jonsson Comprehensive Cancer Center, University of California at Los Angeles

E

Edward B. Garon

S

Samuel W. French

Department of Pathology and Laboratory Medicine, David Geffen School of Medicine, University of California at Los Angeles

C

Clara E. Magyar

Department of Pathology and Laboratory Medicine, David Geffen School of Medicine, University of California at Los Angeles

S

Sarah M. Dry

Department of Pathology and Laboratory Medicine, David Geffen School of Medicine, University of California at Los Angeles

C

Clara M. Lajonchere

Jonsson Comprehensive Cancer Center, University of California at Los Angeles

D

Daniel Geschwind

G

Gina Choi

Department of Medicine, David Geffen School of Medicine, University of California at Los Angeles

S

Sammy Saab

Department of Medicine, David Geffen School of Medicine, University of California at Los Angeles

A

Akshay Shetty

Department of Medicine, David Geffen School of Medicine, University of California at Los Angeles

C

Carrie R. Wong

Vatche and Tamar Manoukian Division of Digestive Diseases, Department of Medicine, University of California

K

Kevin G. King

Department of Radiological Sciences, David Geffen School of Medicine, University of California at Los Angeles

D

David S. Lu

Department of Radiological Sciences, David Geffen School of Medicine, University of California at Los Angeles

S

Steven S. Raman

Department of Radiological Sciences, David Geffen School of Medicine, University of California at Los Angeles

X

Xiyan Xiang

K

Kirti Shetty

Division of Gastroenterology and Hepatology, School of Medicine, University of Maryland

L

Lopa Mishra

S

Sanaz Memarzadeh

Jonsson Comprehensive Cancer Center, University of California at Los Angeles

Y

Yan Liu

F

Frank Alber

W

William Hsu

Department of Radiological Sciences, David Geffen School of Medicine, University of California at Los Angeles

K

Kostyantyn Krysan

Department of Medicine, David Geffen School of Medicine, University of California at Los Angeles

S

Steven M. Dubinett

D

Denise R. Aberle

Department of Radiological Sciences, David Geffen School of Medicine, University of California at Los Angeles

V

Vatche Agopian

Department of Surgery, David Geffen School of Medicine, University of California at Los Angeles

S

Steven-Huy B. Han

Department of Medicine, David Geffen School of Medicine, University of California at Los Angeles

W

Wing Hung Wong

Department of Statistics, Stanford University

X

Xiaohui Ni

EarlyDiagnostics Inc.

W

Wenyuan Li

Department of Biochemistry, The University of Texas Southwestern Medical Center, 5323 Harry Hines Blvd., Dallas, Texas 75390, United States

X

Xianghong Jasmine Zhou

Department of Pathology and Laboratory Medicine, David Geffen School of Medicine, University of California at Los Angeles