Journal: Analytical chemistry
This study presents the development of a novel, highly sensitive methylation detection method for fragmented circulating cell-free DNA (cfDNA) called dRE-STEM. This method combines double restriction enzyme digestion with specific terminal-mediated PCR.
Unlike traditional methods, dRE-STEM does not require bisulfite conversion, which simplifies the process.
It was validated in a colorectal cancer case-control study, with the following results:
- Detection of methylation ratios as low as 3% from only 4 ng of cfDNA
- Diagnostic sensitivity of 80.2%
- Diagnostic specificity of 80.9%
- Performance superior to conventional protein biomarkers
The dRE-STEM method shows promise for accurate methylation quantification in low-abundance, fragmented cfDNA, supporting its potential use in routine clinical cancer diagnostics.