A 5-hydroxymethylcytosine detection method based on triple cascade signal amplification
By employing a triple-cascaded signal amplification detection method, combined with specific glycosylation, magnetic separation, and enzymatic amplification techniques, the sensitivity and specificity issues of 5hmC detection have been resolved, achieving efficient and accurate detection of 5hmC, which is suitable for clinical sample analysis.
CN122326720APending Publication Date: 2026-07-03QINGDAO UNIV OF SCI & TECH
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Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- QINGDAO UNIV OF SCI & TECH
- Filing Date
- 2026-06-05
- Publication Date
- 2026-07-03
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Abstract
This invention discloses a method for detecting 5-hydroxymethylcytosine based on triple cascade signal amplification, belonging to the field of biochemical detection technology. The method includes the following steps: 1) preparation of 5hmC-specific glycosylation and azide labeling; 2) magnetic bead capture and enrichment separation of target DNA; 3) TdT-mediated nucleic acid end polymerization and extension; 4) APE1-triggered primer release and rolling circle amplification; 5) DNase-catalyzed cleavage and fluorescence signal detection. This invention is bisulfite-free and sequence-independent, achieving extremely low background and ultra-high sensitivity detection with a detection limit of 4.41 fM. It can be widely applied to early clinical cancer screening, biomarker detection for neurological or cardiovascular diseases, and provides a powerful platform for low-abundance epigenetic analysis in clinical diagnosis and biomedical research.
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