SPMSP: A Solid-phase PCR with Colorimetric Read-out for Quantitative Methylation Analysis

Abstract

Background: As DNA methylation has been determined as an important diagnostic biomarker in cancer management, methods for the detection of quantitative DNA methylation which are high-throughput and low-cost are needed. Materials and Methods: In this work, we introduce the solid-phase methylation-specific PCR (SPMSP) as a method for quantitative methylation analysis. SPMSP combines methylation-specific DNA amplification on a solid phase with subsequent colorimetric detection in an ELISA-based format. Results: In contrast to existing methods for quantitative methylation analysis, SPMSP can be carried out with standard laboratory equipment, i.e. a thermocycler and an ELISA reader. With this method, DNA methylation in the promoter of the APC tumour suppressor gene was quantified in a set of colorectal carcinoma samples. Conclusion: As SPMSP can be modified to the analysis of other promoter sequences and is also adaptable to a high-throughput system, SPMSP offers a platform for methylation profiling with widespread applications in cancer research and management.