Department of Chemistry, University of Washington, Seattle, Washington 98195, USA.
Lab Chip. 2019 Mar 13;19(6):1035-1040. doi: 10.1039/c8lc01223g.
Digital nucleic acid amplification and detection methods provide excellent sensitivity and specificity and allow absolute quantification of target nucleic acids. Isothermal methods such as digital loop-mediated isothermal amplification (digital LAMP) have potential for use in rapid disease diagnosis in low-resource settings due to their speed and lack of thermal cycling. We previously developed a self-digitization (SD) chip, a simple microfluidics device that automatically digitizes a sample into an array of nanoliter wells, for use in digital LAMP. In this work, we improve the SD chip design to increase sample loading efficiency, speed, and completeness, and test a range of well volumes and numbers. We demonstrate the diagnostic capability of this platform by applying it to quantifying human papillomavirus 18 gene.
数字核酸扩增和检测方法提供了极好的灵敏度和特异性，并允许对靶核酸进行绝对定量。等温方法，如数字环介导等温扩增（digital LAMP），由于其速度和无需热循环，有可能用于资源匮乏环境中的快速疾病诊断。我们之前开发了一种自数字化（SD）芯片，这是一种简单的微流控装置，可将样品自动数字化成纳米升孔阵列，用于数字 LAMP。在这项工作中，我们改进了 SD 芯片设计，以提高样品加载效率、速度和完整性，并测试了一系列不同的孔体积和数量。我们通过应用该平台定量检测人乳头瘤病毒 18 基因来证明该平台的诊断能力。
