Multi-STEM MePCR: a bisulfite-free, multiplex, highly sensitive and highly specific assay to measure DNA methylation.

ABSTRACT: Aberrant CpG island methylation serves as a pivotal biomarker for cancer diagnosis, with accuracy substantially enhanced by analyzing multiple loci. Current techniques, such as bisulfite conversion or restriction enzyme-based methods, often fall short in delivering efficient multiplexed genomic methylation analysis using standard PCR platforms. Herein, we introduce an innovative bisulfite-free, multiplex assay-multiple specific terminal mediated methylation PCR (multi-STEM MePCR). This assay integrates a methylation-dependent restriction endonuclease (MDRE) with a novel multiplex PCR, leveraging innovative stem-loop structured assays for simultaneous detection of multiple CpG sites. As a proof-of-concept, the multi-STEM MePCR platform simultaneously achieved quantification of three methyla

SUBMITTER: Yang H 

PROVIDER: S-EPMC12178177 | biostudies-literature | 2025 Jul

REPOSITORIES: biostudies-literature