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Peer-reviewedCC BYBMC gastroenterology

Methylation-sensitive restriction endonuclease-based amplification assay for detecting DNA methylation in colorectal cancer: a cross-sectional study.

  • Jiali Wu1
  • Zhikang Li2
  • Lu Ai1
  • Anzhi Huang3
  • Shayuanzi Huang4
  • Ruizhao Zhang5
  • Guoshan He5
  • Meifang He6
  • Ruizhi Wang7,8
  • Dong Wang9,10,11
11 affiliations
  1. 1Department of Laboratory Medicine, The First Affiliated Hospital of Sun Yat-sen University, 58 Zhongshan road II, Guangzhou, Guangdong, China.
  2. 2Laboratory of General Surgery, The First Affiliated Hospital of Sun Yat-Sen University, 58 Zhongshan road II, Guangzhou, Guangdong, China.
  3. 3Guangdong Cardiovascular Institute, Medical Research Institute, School of Basic Medical Science, Guangdong Provincial People's Hospital, Guangzhou, Guangdong, China.
  4. 4Department of Laboratory Medicine, Guangxi Hospital Division of The First Affiliated Hospital of Sun Yat-sen University, Nanning, Guangxi, China.
  5. 5Guangzhou Quality Testing & Inspection Institute, Guangzhou, Guangdong, China.
  6. 6Laboratory of General Surgery, The First Affiliated Hospital of Sun Yat-Sen University, 58 Zhongshan road II, Guangzhou, Guangdong, China. [email protected].
  7. 7Department of Laboratory Medicine, The First Affiliated Hospital of Sun Yat-sen University, 58 Zhongshan road II, Guangzhou, Guangdong, China. [email protected].
  8. 8Advanced Medical Technology Center, The First Affiliated Hospital, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, Guangdong, China. [email protected].
  9. 9Department of Laboratory Medicine, The First Affiliated Hospital of Sun Yat-sen University, 58 Zhongshan road II, Guangzhou, Guangdong, China. [email protected].
  10. 10Department of Laboratory Medicine, Guangxi Hospital Division of The First Affiliated Hospital of Sun Yat-sen University, Nanning, Guangxi, China. [email protected].
  11. 11Guangdong Provincial Clinical Research Center for Laboratory Medicine, Guangzhou, P. R. China. [email protected].
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Abstract

Background

This study evaluates the clinical utility of combined SDC2 and ADHFE1 methylation analysis for colorectal cancer (CRC) detection using a methylation-sensitive restriction endonuclease (MSRE)-based assay.Methods

Leveraging the specificity of MSRE, we developed a streamlined amplification assay that directly analyzes methylation status of SDC2 and ADHFE1 after mixing nucleic acids with PCR reagent. A cohort of 274 participants was stratified into six groups: CRC (n = 117), advanced adenoma (AA, n = 22), polyps (n = 30), intestinal disease control (IDC, n = 39), other gastrointestinal disease control (OGDC, n = 24), and healthy controls (HC, n = 42). SDC2 and ADHFE1 methylation levels in fecal samples were assessed, and diagnostic performance was evaluated using receiver operating characteristic (ROC) curve analysis.Results

The CRC group exhibited significantly higher SDC2 and ADHFE1 methylation positivity rates compared to all comparator groups, including AA, polyps, IDC, OGDC, and HC (P < 0.01). No significant differences were observed in the methylation positivity rates of SDC2, ADHFE1 or their combination across different TNM stages and levels of tumor differentiation. Joint SDC2 and ADHFE1 analysis increased sensitivity for CRC detection to 82.9% (vs. 64.1% and 65.0% for ADHFE1 and SDC2) with an AUC of 0.865 (95% CI: 0.817-0.913). For early-stage CRC (stages I + II), the dual-gene panel achieved an AUC of 0.872. However, the ability to discriminate between AA and HC (AUC: 0.693) or IDC (AUC: 0.615) remained limited in our dual-gene assay.Conclusions

The combined SDC2/ADHFE1 MSRE-based assay significantly improves diagnostic sensitivity for CRC, demonstrating superior differentiation of CRC from benign intestinal pathologies and precancerous lesions (AA and polyps). This non-invasive approach shows promise as a supplementary tool for CRC screening.

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Cite

BibTeXRIS
BibTeX
@article{wu2026methylationsensitive,
  title = {{Methylation-sensitive restriction endonuclease-based amplification assay for detecting DNA methylation in colorectal cancer: a cross-sectional study.}},
  author = {Jiali Wu and Zhikang Li and Lu Ai and Anzhi Huang and Shayuanzi Huang and Ruizhao Zhang and Guoshan He and Meifang He and Ruizhi Wang and Dong Wang},
  journal = {BMC gastroenterology},
  year = {2026},
  month = aug,
  doi = {10.1186/s12876-026-05034-7},
  url = {https://europepmc.org/article/MED/42958101},
  note = {CC BY},
}
RIS
TY  - JOUR
TI  - Methylation-sensitive restriction endonuclease-based amplification assay for detecting DNA methylation in colorectal cancer: a cross-sectional study.
AU  - Jiali Wu
AU  - Zhikang Li
AU  - Lu Ai
AU  - Anzhi Huang
AU  - Shayuanzi Huang
AU  - Ruizhao Zhang
AU  - Guoshan He
AU  - Meifang He
AU  - Ruizhi Wang
AU  - Dong Wang
PY  - 2026
DA  - 2026/08/03
JO  - BMC gastroenterology
DO  - 10.1186/s12876-026-05034-7
UR  - https://europepmc.org/article/MED/42958101
AB  - Background  This study evaluates the clinical utility of combined SDC2 and ADHFE1 methylation analysis for colorectal cancer (CRC) detection using a methylation-sensitive restriction endonuclease (MSRE)-based assay.Methods  Leveraging the specificity of MSRE, we developed a streamlined amplification assay that directly analyzes methylation status of SDC2 and ADHFE1 after mixing nucleic acids with PCR reagent. A cohort of 274 participants was stratified into six groups: CRC (n = 117), advanced adenoma (AA, n = 22), polyps (n = 30), intestinal disease control (IDC, n = 39), other gastrointestinal disease control (OGDC, n = 24), and healthy controls (HC, n = 42). SDC2 and ADHFE1 methylation levels in fecal samples were assessed, and diagnostic performance was evaluated using receiver operating characteristic (ROC) curve analysis.Results  The CRC group exhibited significantly higher SDC2 and ADHFE1 methylation positivity rates compared to all comparator groups, including AA, polyps, IDC, OGDC, and HC (P < 0.01). No significant differences were observed in the methylation positivity rates of SDC2, ADHFE1 or their combination across different TNM stages and levels of tumor differentiation. Joint SDC2 and ADHFE1 analysis increased sensitivity for CRC detection to 82.9% (vs. 64.1% and 65.0% for ADHFE1 and SDC2) with an AUC of 0.865 (95% CI: 0.817-0.913). For early-stage CRC (stages I + II), the dual-gene panel achieved an AUC of 0.872. However, the ability to discriminate between AA and HC (AUC: 0.693) or IDC (AUC: 0.615) remained limited in our dual-gene assay.Conclusions  The combined SDC2/ADHFE1 MSRE-based assay significantly improves diagnostic sensitivity for CRC, demonstrating superior differentiation of CRC from benign intestinal pathologies and precancerous lesions (AA and polyps). This non-invasive approach shows promise as a supplementary tool for CRC screening.
SN  - 1471-230X
ER  -