An Automated Multi-Sample Digital LAMP Platform for Quantitative Detection of SDC2 Methylation in Colorectal Cancer

Abstract

DNA methylation of the SDC2 gene has emerged as a critical biomarker for early detection of colorectal cancer (CRC). However, effective clinical application demands analytical platforms that offer enhanced accuracy, speed, and sample high-throughput capacity. In this study, we present an automated multisample digital loop-mediated isothermal amplification (dLAMP) platform designed to meet these requirements. Our platform integrates a dual-volume microfluidic chip capable of analyzing 4 or 8 samples in parallel, significantly enhancing sample throughput and efficiency. Integrated with automatic sample injection and advanced image processing, the platform achieves streamlined, multistep analysis of SDC2 methylation levels. We designed methylation-specific (MS) and unmethylation-specific (UMS) LAMP primers with high specificity. Validation using CRC cell lines (HCT116 and Caco-2) and clinical tissue samples demonstrated robust performance with high sensitivity, specificity and reproducibility. The results showed elevated SDC2 methylation was detected in tumor tissues across all CRC stages. This integrated automated dLAMP platform provides a robust, multisample high-throughput tool for methylation-based diagnostics and holds strong potential for noninvasive early cancer screening and clinical biomarker applications.