Detection of MGMT methylation status using a Lab-on-Chip compatible isothermal amplification method.

Abstract

The growing cancer burden necessitates the development of cost-effective solutions that provide rapid, precise and personalised information to improve patient outcome. The aim of this study was to develop a novel, Lab-on-Chip compatible method for the detection and quantification of DNA methylation for MGMT, a well-established molecular biomarker for glioblastoma, with direct clinical translation as a predictive target. A Lab-on-Chip compatible isothermal amplification method (LAMP) was used to test its efficacy for detection of sequence-specific methylated regions of MGMT, with the method's specificity and sensitivity to have been compared against gold-standards (MethyLight, JumpStart). Our LAMP primer combinations were shown to be specific to the MGMT methylated region, while sensitivity assays determined that the amplification methods were capable of running at clinically relevant DNA concentrations of 0.2 - 20 ng/µL. For the first time, the ability to detect the presence of DNA methylation on bisulfite converted DNA was demonstrated on a Lab-on-Chip setup, laying the foundation for future applications of this platform to other epigenetic biomarkers in a point-of-care setting.