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Peer-reviewedCC BYCancers

A Dual-Marker DNA Methylation Assay Enables High-Sensitivity Detection of Malignant Effusions Across Cancer Types.

  • Ting Wang1
  • Ting Liang2
  • Matthew Kafarski3
  • Jian Liu4
  • Yi Jing2
  • Wen Liao2
  • Hang Xing5
  • Jia Feng2
  • Tingting Bian2
  • Lei Liu2
  • Hui Sun2
  • Xiaoli Zheng1
9 more authors
  • Bin She6
  • Xiaosha Ren6
  • Joanne Lee7
  • Qiushi Lin3
  • Ming Chen3
  • Liang Cheng7
  • Hong Ge1
  • Xiaoqun Dong3
  • Yifei Liu2
7 affiliations
  1. 1Department of Radiation Oncology, The Affiliated Cancer Hospital of Zhengzhou University & Henan Cancer Hospital, No. 127 Dongming Road, Jinshui District, Zhengzhou 450008, China.
  2. 2Department of Pathology, Affiliated Hospital of Nantong University, Medical School of Nantong University, Nantong 226001, China.
  3. 3Precision Health Program, Department of Radiology, College of Human Medicine, Michigan State University, East Lansing, MI 48824, USA.
  4. 4Department of Oncology, Affiliated Hospital of Nantong University, Medical School of Nantong University, No. 20 Xisi Road, Chongchuan District, Nantong 226001, China.
  5. 5Rhode Island Hospital, Alpert Medical School of Brown University, Providence, RI 02912, USA.
  6. 6Director of Pathology Business Unit, Shanghai ZJ Bio-Tech CO., Ltd., Building 9, No. 2168 Chenhang Hwy, Shanghai 201114, China.
  7. 7Department of Pathology and Laboratory Medicine, Brown University Warren Alpert Medical School, 142 Boren Street, Providence, RI 02912, USA.
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Abstract

Objective: This study evaluated the diagnostic value of a combined SHOX2 and RASSF1A methylation assay (LungMe®) in differentiating malignant from benign pleural and peritoneal effusions. Methods: A total of 696 patients with pleural or peritoneal effusions were prospectively enrolled. Effusion sediment samples were analyzed by liquid-based cytology (TCT), cell block histology, and methylation-specific PCR for both SHOX2 and RASSF1A. The final diagnosis was confirmed by clinical, imaging, and pathological follow-up. Results: Lung cancer was the predominant cause of malignant pleural effusions (71.96%), while malignant peritoneal effusions mainly resulted from ovarian, gastric, and liver cancers. The optimal ΔCt cutoffs for SHOX2 and RASSF1A were ΔCt ≤ 9 and ΔCt ≤ 12, respectively. The combined methylation assay achieved a sensitivity of 79.1% and specificity of 90.4% for malignant effusions. When combined with cytology, the overall sensitivity increased from 34.7% (TCT alone) to 91.7%, with a positive predictive value (PPV) of 93.9% and a negative predictive value (NPV) of 86.7%. The assay showed high sensitivity across multiple tumor types, including 100% for esophageal and breast cancers, and ≥80% for lung cancer, gastric cancer, lymphoma, and cholangiocarcinoma. Conclusions: The combined SHOX2 and RASSF1A methylation assay has high diagnostic value for benign-malignant discrimination of pleural and peritoneal effusions, improving diagnostic sensitivity when combined with conventional cytology. Thus, within the context of this single-center study, the LungMe® assay demonstrates promise in expanding pan-cancer diagnostics, but will require further multicenter external validation and prospective clinical studies before routine clinical adoption.

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Cite

BibTeXRIS
BibTeX
@article{wang2026dualmarker,
  title = {{A Dual-Marker DNA Methylation Assay Enables High-Sensitivity Detection of Malignant Effusions Across Cancer Types.}},
  author = {Ting Wang and Ting Liang and Matthew Kafarski and Jian Liu and Yi Jing and Wen Liao and Hang Xing and Jia Feng and Tingting Bian and Lei Liu and Hui Sun and Xiaoli Zheng and Bin She and Xiaosha Ren and Joanne Lee and Qiushi Lin and Ming Chen and Liang Cheng and Hong Ge and Xiaoqun Dong and Yifei Liu},
  journal = {Cancers},
  year = {2026},
  month = oct,
  doi = {10.3390/cancers18193176},
  url = {https://europepmc.org/article/MED/42882870},
  note = {CC BY},
}
RIS
TY  - JOUR
TI  - A Dual-Marker DNA Methylation Assay Enables High-Sensitivity Detection of Malignant Effusions Across Cancer Types.
AU  - Ting Wang
AU  - Ting Liang
AU  - Matthew Kafarski
AU  - Jian Liu
AU  - Yi Jing
AU  - Wen Liao
AU  - Hang Xing
AU  - Jia Feng
AU  - Tingting Bian
AU  - Lei Liu
AU  - Hui Sun
AU  - Xiaoli Zheng
AU  - Bin She
AU  - Xiaosha Ren
AU  - Joanne Lee
AU  - Qiushi Lin
AU  - Ming Chen
AU  - Liang Cheng
AU  - Hong Ge
AU  - Xiaoqun Dong
AU  - Yifei Liu
PY  - 2026
DA  - 2026/10/01
JO  - Cancers
DO  - 10.3390/cancers18193176
UR  - https://europepmc.org/article/MED/42882870
AB  - Objective: This study evaluated the diagnostic value of a combined SHOX2 and RASSF1A methylation assay (LungMe®) in differentiating malignant from benign pleural and peritoneal effusions. Methods: A total of 696 patients with pleural or peritoneal effusions were prospectively enrolled. Effusion sediment samples were analyzed by liquid-based cytology (TCT), cell block histology, and methylation-specific PCR for both SHOX2 and RASSF1A. The final diagnosis was confirmed by clinical, imaging, and pathological follow-up. Results: Lung cancer was the predominant cause of malignant pleural effusions (71.96%), while malignant peritoneal effusions mainly resulted from ovarian, gastric, and liver cancers. The optimal ΔCt cutoffs for SHOX2 and RASSF1A were ΔCt ≤ 9 and ΔCt ≤ 12, respectively. The combined methylation assay achieved a sensitivity of 79.1% and specificity of 90.4% for malignant effusions. When combined with cytology, the overall sensitivity increased from 34.7% (TCT alone) to 91.7%, with a positive predictive value (PPV) of 93.9% and a negative predictive value (NPV) of 86.7%. The assay showed high sensitivity across multiple tumor types, including 100% for esophageal and breast cancers, and ≥80% for lung cancer, gastric cancer, lymphoma, and cholangiocarcinoma. Conclusions: The combined SHOX2 and RASSF1A methylation assay has high diagnostic value for benign-malignant discrimination of pleural and peritoneal effusions, improving diagnostic sensitivity when combined with conventional cytology. Thus, within the context of this single-center study, the LungMe® assay demonstrates promise in expanding pan-cancer diagnostics, but will require further multicenter external validation and prospective clinical studies before routine clinical adoption.
SN  - 2072-6694
ER  -