Shahriari Shadi, Selvaganapathy P Ravi
Department of Mechanical Engineering, McMaster University, Hamilton, ON L8S 4L8, Canada.
School of Biomedical Engineering, McMaster University, Hamilton, ON L8S 4L8, Canada.
Gels. 2024 Mar 8;10(3):186. doi: 10.3390/gels10030186.
Sepsis, a life-threatening condition resulting from a failing host response to infection, causes millions of deaths annually, necessitating rapid and simple prognostic assessments. A variety of genomic and proteomic biomarkers have been developed for sepsis. For example, it has been shown that the level of plasma cell-free DNA (cfDNA) and circulating histones increases considerably during sepsis, and they are linked with sepsis severity and mortality. Developing a diagnostic tool that is capable of assessing such diverse biomarkers is challenging as the detection methodology is quite different for each. Here, a fully integrated microfluidic device capable of detecting a genomic biomarker (cfDNA) and a proteomic biomarker (total circulating histones) using a common detection platform has been demonstrated. The microfluidic device utilizes dehydrated agarose gates loaded with pH-specific agarose to electrophoretically trap cfDNA and histones at their respective isoelectric points. It also incorporates fluorescent dyes within the device, eliminating the need for off-chip sample preparation and allowing the direct testing of plasma samples without the need for labeling DNA and histones with fluorescent dyes beforehand. Xurography, which is a low-cost and rapid method for fabrication of microfluidics, is used in all the fabrication steps. Experimental results demonstrate the effective accumulation and separation of cfDNA and histones in the agarose gates in a total processing time of 20 min, employing 10 and 30 Volts for cfDNA and histone accumulation and detection, respectively. The device can potentially be used to distinguish between the survivors and non-survivors of sepsis. The integration of the detection of both biomarkers into a single device and dye immobilization enhances its clinical utility for rapid point-of-care assessment of sepsis prognosis.
脓毒症是宿主对感染的反应失败所导致的一种危及生命的病症，每年造成数百万人死亡，因此需要快速且简便的预后评估方法。针对脓毒症，已经开发出了多种基因组和蛋白质组生物标志物。例如，研究表明，在脓毒症期间，血浆游离DNA（cfDNA）和循环组蛋白的水平会大幅升高，且它们与脓毒症的严重程度和死亡率相关。开发一种能够评估如此多样生物标志物的诊断工具具有挑战性，因为每种生物标志物的检测方法差异很大。在此，已展示了一种完全集成的微流控装置，该装置能够使用通用检测平台检测基因组生物标志物（cfDNA）和蛋白质组生物标志物（总循环组蛋白）。该微流控装置利用装载有特定pH值琼脂糖的脱水琼脂糖门，在其各自的等电点处通过电泳捕获cfDNA和组蛋白。它还在装置内整合了荧光染料，无需进行芯片外样品制备，并且无需事先用荧光染料标记DNA和组蛋白即可直接检测血浆样品。在所有制造步骤中都使用了Xurography，这是一种用于制造微流控器件的低成本且快速的方法。实验结果表明，在总共20分钟的处理时间内，cfDNA和组蛋白在琼脂糖门中实现了有效积累和分离，cfDNA和组蛋白的积累及检测分别采用10伏和30伏电压。该装置有可能用于区分脓毒症的幸存者和非幸存者。将两种生物标志物的检测集成到单个装置中以及染料固定化增强了其在脓毒症预后快速即时护理评估中的临床实用性。
