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LAMP-MS for Locus-Specific Visual Quantification of DNA 5 mC and RNA m6A Using Ultra-Low Input
Runyu Xie1, Xiaotong Yang1, Weizhi He1
1Shanghai Cancer Center, Institutes of Biomedical Sciences, Shanghai Key Laboratory of Medical Epigenetics, International Co-laboratory of Medical Epigenetics and Metabolism (Ministry of Science and Technology), Shanghai Medical College of Fudan University, Shanghai, China, 200032.
Angewandte Chemie (International Ed. in English)
|November 3, 2024
Summary
A new LAMP-MS technology enhances circulating cell-free DNA (cfDNA) analysis for disease screening. This method offers sensitive, cost-effective detection of cfDNA methylation markers for improved diagnostics.
Area of Science:
- Biotechnology
- Molecular Diagnostics
- Genomics
Background:
- Circulating cell-free DNA (cfDNA) holds promise for non-invasive disease screening.
- Current methods for cfDNA analysis face challenges in sensitivity, specificity, and cost-efficiency.
- Detecting cfDNA methylation patterns is crucial for early disease detection, particularly for colorectal cancer (CRC).
Purpose of the Study:
- To introduce and validate LAMP-MS, a novel technology for sensitive and quantitative cfDNA methylation analysis.
- To overcome limitations of existing cfDNA screening methods.
- To demonstrate the potential of LAMP-MS for clinical applications in disease detection.
Main Methods:
- Development of LAMP-MS, integrating linear amplification with single-base quantitative nucleic acid mass spectrometry on silicon chips.
- Application of LAMP-MS for unbiased amplification of cfDNA (as little as 1 ng).
- Site-specific quantification of multiple 5-methylcytosine (5mC) sites in cfDNA and validation using DNA probes and patient plasma.
Main Results:
- LAMP-MS enables accurate, high-resolution quantification of cfDNA methylation markers.
- The technology demonstrated accuracy validated by mass spectrometric peak areas.
- Expanded capability of Mass Array technology to quantify RNA m6A modification sites was shown.
Conclusions:
- LAMP-MS is a promising technology for sensitive and cost-effective cfDNA methylation detection.
- The method's ability to work with ultra-low input material and provide visually interpretable output makes it suitable for clinical settings.
- LAMP-MS offers a robust platform for nucleic acid methylation detection and quantification in diagnostics.
