甲基化
DNA甲基化
结直肠癌
生物标志物
吞吐量
化学
数字聚合酶链反应
环介导等温扩增
计算生物学
癌症
熔化曲线分析
癌症研究
样品(材料)
癌症生物标志物
转录组
DNA微阵列
微流控
微阵列
炸薯条
微流控芯片
亚硫酸氢盐测序
DNA
作者
Hao He,Yanzhao Wu,Lai Wei,Lin Zhou,Zhirui Liu,Nan Rong,Chunxiong Luo,Chanqiong Zhang,Wei Yang
标识
DOI:10.1021/acs.analchem.5c03244
摘要
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.
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