C. Eugene Bennett Department of Chemistry, West Virginia University, Morgantown, WV 26506, USA.
Department of Mechanical and Aerospace Engineering, West Virginia University, Morgantown, WV 26506, USA.
Sensors (Basel). 2024 Jun 30;24(13):4266. doi: 10.3390/s24134266.
Nucleic acid tests are key tools for the detection and diagnosis of many diseases. In many cases, the amplification of the nucleic acids is required to reach a detectable level. To make nucleic acid amplification tests more accessible to a point-of-care (POC) setting, isothermal amplification can be performed with a simple heating source. Although these tests are being performed in bulk reactions, the quantification is not as accurate as it would be with digital amplification. Here, we introduce the use of the vibrating sharp-tip capillary for a simple and portable system for tunable on-demand droplet generation. Because of the large range of droplet sizes possible and the tunability of the vibrating sharp-tip capillary, a high dynamic range (~2 to 6000 copies/µL) digital droplet loop-mediated isothermal amplification (ddLAMP) system has been developed. It was also noted that by changing the type of capillary on the vibrating sharp-tip capillary, the same mechanism can be used for simple and portable DNA fragmentation. With the incorporation of these elements, the present work paves the way for achieving digital nucleic acid tests in a POC setting with limited resources.
核酸检测是许多疾病检测和诊断的关键工具。在许多情况下，需要扩增核酸以达到可检测的水平。为了使核酸扩增检测更适用于即时检测（POC）环境，可以使用简单的加热源进行等温扩增。尽管这些测试是在批量反应中进行的，但定量的准确性不如数字扩增。在这里，我们介绍了使用振动式尖锐尖端毛细管来构建简单便携的可调按需液滴生成系统。由于可能产生的液滴尺寸范围很大且振动式尖锐尖端毛细管的可调性，因此已经开发出了具有高动态范围（~2 到 6000 拷贝/µL）的数字液滴环介导等温扩增（ddLAMP）系统。还注意到，通过改变振动式尖锐尖端毛细管上的毛细管类型，可以使用相同的机制进行简单便携的 DNA 片段化。通过纳入这些元素，本工作为在资源有限的即时检测环境中实现数字核酸检测铺平了道路。
