Lamb wave-based molecular diagnosis using DNA hydrogel formation by rolling circle amplification (RCA) process
Lamb wave-based molecular diagnosis using DNA hydrogel formation by rolling circle amplification (RCA) process
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DOI：
10.1016/j.bios.2019.111496
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发表时间：
2019-10-01
影响因子：
12.6
通讯作者：
Lim, Chae Seung
中科院分区：
文献类型：
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作者：
Nam, Jeonghun;Jang, Woong Sik;Lim, Chae Seung
微流体技术的最新发展使基于芯片实验室的分子诊断成为可能。快速准确地诊断传染病对于预防疾病传播至关重要。在这里，我们使用各种参数，包括接触角和粘度的样品液滴，施加的电压，和温度增加的兰姆波为基础的设备的特征。此外，我们证明了兰姆波为基础的设备在临床应用中的功能。滚环扩增（RCA）过程的最佳温度为30 ° C，并且其通过在17V下产生兰姆波来实现。由于RCA过程的基因扩增可以通过由于样品液滴中DNA水凝胶形成而导致的粘度增加来检测，这引起悬浮颗粒的声流速度降低。在我们的兰姆波为基础的设备，等温扩增的目标核酸可以成功地检测在30分钟内使用10 μ L的固着液滴，并通过比较商业实时荧光分析验证。我们的设备能够实现简单和低成本的分子诊断，可应用于资源有限的临床环境。
Recent developments in microfluidics enable the lab-on-a-chip-based molecular diagnosis. Rapid and accurate diagnosis of infectious diseases is critical for preventing the transmission of the disease. Here, we characterize a Lamb wave-based device using various parameters including the contact angle and viscosity of the sample droplet, the applied voltage, and the temperature increase. Additionally, we demonstrate the functionality of the Lamb wave-based device in clinical application. Optimal temperature for rolling circle amplification (RCA) process is 30 degrees C, and it was achieved by Lamb wave generation at 17 V. Gene amplification due to RCA process could be detected by viscosity increase due to DNA hydrogel formation in a sample droplet, which induced the acoustic streaming velocity of suspended particles to be decreased. In our Lamb wave-based device, isothermal amplification of target nucleic acids could be successfully detected within 30 min using 10 mu L of sessile droplet, and was validated by comparing that of commercial real-time fluorescence analysis. Our device enables simple and low-cost molecular diagnosis, which can be applied to resource-limited clinical settings.