Park Juhwan, Han Dong Hyun, Hwang Sang-Hyun, Park Je-Kyun
Department of Bio and Brain Engineering, Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Yuseong-gu, Daejeon 34141, Republic of Korea.
Lab Chip. 2020 Sep 21;20(18):3346-3353. doi: 10.1039/d0lc00432d. Epub 2020 Jul 6.
Molecular diagnostics can provide a powerful diagnostic tool since it can detect pathogens with high sensitivity, but complicated sample preparation procedures limit its widespread use as an on-site detection tool that relies on the skilled person and external equipment. To resolve these limitations, we report a solid-phase nucleic acid purification using a finger-actuated microfluidic device, which can control a set amount of flow regardless of differences in end-users. To increase the recovery rate, a finger-actuated reciprocator was newly developed and integrated into the microfluidic device that can efficiently react with silica microbeads and reagents. After verifying the finger-actuated microfluidic reciprocator, the effect of the reciprocating flow on the recovery rate was assessed to purify the standard DNA of the hepatitis B virus (HBV). The recovery rate was increased up to ∼50% and 955 to 955 000 IU mL of HBV standard DNA was successfully purified and detected by a real-time polymerase chain reaction. Furthermore, the proposed microfluidic device was exploited to purify the HBV DNA from the patient's blood plasma samples.
分子诊断可以提供一种强大的诊断工具,因为它能够高灵敏度地检测病原体,但复杂的样品制备程序限制了其作为一种依赖技术人员和外部设备的现场检测工具的广泛应用。为了解决这些限制,我们报告了一种使用手指驱动微流控装置的固相核酸纯化方法,该装置可以控制设定的流量,而不受最终用户差异的影响。为了提高回收率,新开发了一种手指驱动的往复装置并将其集成到微流控装置中,该装置可以与二氧化硅微珠和试剂高效反应。在验证了手指驱动的微流控往复装置后,评估了往复流动对回收率的影响,以纯化乙型肝炎病毒(HBV)的标准DNA。回收率提高到约50%,通过实时聚合酶链反应成功纯化并检测到955至955000 IU/mL的HBV标准DNA。此外,所提出的微流控装置被用于从患者血浆样本中纯化HBV DNA。