Xu Zhiheng, Yin Kun, Ding Xiong, Li Ziyue, Sun Xuanhao, Li Baikun, Lalla Rajesh V, Gross Robert, Liu Changchun
Department of Biomedical Engineering, University of Connecticut Health Center, Farmington, CT, 06030, USA.
Department of Physiology and Neurobiology, University of Connecticut, Storrs, CT, 06269, USA.
Biosens Bioelectron. 2021 Aug 15;186:113306. doi: 10.1016/j.bios.2021.113306. Epub 2021 May 7.
A simple, disposable, and integrated electronic-tube cap (E-tube cap) for DNA detection at the point-of-care was designed, fabricated, and tested. The E-tube cap contains a 3D printed electrode substrate for DNA extraction and label-free pH sensing detection. One Flinders Technology Associates (Whatman FTA) membrane was incorporated into the 3D printed electrode substrate for the isolation, concentration, and purification of DNA. The E-tube cap with captured DNA by the membrane was inserted directly into a reaction tube for loop-mediated isothermal amplification (LAMP). The isothermal amplification process was monitored in real-time by a 3D printed electrochemical electrode coated with pH-sensitive material (carbon/iridium oxide layer). The pH sensing electrode showed an excellent linear response within the pH range of 6-9 with a slope of -31.32 ± 0.5 mV/pH at room temperature. The utility of the integrated E-tube cap was demonstrated by detecting the presence of lambda DNA spiked in saliva samples with a sensitivity of 100 copies per mL sample within 30 min. Such a simple, rapid, and affordable diagnostic device is particularly suitable for point-of-care molecular diagnostics of infectious diseases.
设计、制造并测试了一种用于即时检测DNA的简单、一次性且集成的电子管帽(E管帽)。该E管帽包含一个用于DNA提取和无标记pH传感检测的3D打印电极基板。将一张弗林德斯技术协会(Whatman FTA)膜并入3D打印电极基板中,用于DNA的分离、浓缩和纯化。将通过膜捕获了DNA的E管帽直接插入反应管中进行环介导等温扩增(LAMP)。通过涂有pH敏感材料(碳/氧化铱层)的3D打印电化学电极实时监测等温扩增过程。在室温下,pH传感电极在6-9的pH范围内显示出优异的线性响应,斜率为-31.32±0.5 mV/pH。通过检测唾液样本中添加的λDNA,在30分钟内灵敏度达到每毫升样本100个拷贝,证明了集成E管帽的实用性。这种简单、快速且经济实惠的诊断设备特别适用于传染病的即时分子诊断。
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