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Multiple on-line active valves based centrifugal microfluidics for dynamic solid-phase enrichment and purification of viral nucleic acid
Lab on a Chip ( IF 6.1 ) Pub Date : 2024-05-23 , DOI: 10.1039/d4lc00074a Shunji Li 1 , Chao Wan 1 , Yujin Xiao 1 , Changgen Liu 2 , Xudong Zhao 1 , Ying Zhang 1 , Huijuan Yuan 1 , Liqiang Wu 2 , Chungen Qian 2 , Yiwei Li 1 , Peng Chen 1 , Bi-Feng Liu 1
Lab on a Chip ( IF 6.1 ) Pub Date : 2024-05-23 , DOI: 10.1039/d4lc00074a Shunji Li 1 , Chao Wan 1 , Yujin Xiao 1 , Changgen Liu 2 , Xudong Zhao 1 , Ying Zhang 1 , Huijuan Yuan 1 , Liqiang Wu 2 , Chungen Qian 2 , Yiwei Li 1 , Peng Chen 1 , Bi-Feng Liu 1
Affiliation
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Point of care testing (POCT) of nucleic acids holds significant importance in the realm of infectious disease prevention and control, as well as the advancement of personalized precision medicine. Nevertheless, conventional nucleic acid testing methods continue to face challenges such as prolonged detection times and dependence on extensive specialized equipment and personnel, rendering them unsuitable for point of care applications. Here, we proposed an innovative active centrifugal microfluidic system (ACMS) for automatic nucleic acid extraction, encompassing modules for active valve control and magnetic control. An on-chip centrifugal puncture valve (PV) was devised based on the elastic tolerance differences between silicone membranes and tinfoils to release pre-embedded liquid reagents on demand. Furthermore, we have utilized the returnable valve (RV) technology to accurately control the retention and release of liquids, leveraging the high elastic tolerance of the silicone membrane. By incorporating an online controllable magnetic valve, we have achieved controlled and rapid aggregation and dispersion of magnetic beads. The final chip encapsulates multiple reagents and magnetic beads necessary for nucleic acid extraction. Upon sample addition and loading into the instrument, automated on-chip sample loading and nucleic acid extraction, purification, and collection can be accomplished within 30 minutes, halving the overall operation time and even increasing the efficiency of pseudovirus extraction by three orders of magnitude. Consequently, real-time fluorescence quantitative PCR amplification has successfully detected multiple targets of the SARS-CoV-2 virus (with an impressive detection limit as low as 10 copies per μL), along with targeted sequencing analysis yielding a conformity rate of 99%.
中文翻译：
基于多个在线主动阀的离心微流体用于病毒核酸的动态固相富集和纯化
核酸即时检测（POCT）在传染病预防和控制领域以及个性化精准医疗的发展中具有重要意义。然而，传统的核酸检测方法仍然面临检测时间长、依赖大量专业设备和人员等挑战，使其不适合床旁应用。在这里，我们提出了一种用于自动核酸提取的创新主动离心微流体系统（ACMS），包括主动阀控制和磁控制模块。根据硅胶膜和锡箔之间的弹性公差差异，设计了一种片上离心穿刺阀（PV），可根据需要释放预嵌入的液体试剂。此外，我们还利用可返回阀（RV）技术，利用硅胶膜的高弹性公差来精确控制液体的保留和释放。通过采用在线可控磁阀，我们实现了磁珠的可控、快速聚集和分散。最终的芯片封装了核酸提取所需的多种试剂和磁珠。样品添加并加载到仪器中后，30分钟内即可完成芯片上的自动化样品加载和核酸提取、纯化和收集，使整体操作时间减少一半，甚至将假病毒提取效率提高三个数量级。 因此，实时荧光定量 PCR 扩增已成功检测到 SARS-CoV-2 病毒的多个靶标（检测限低至每 μL 10 个拷贝），并且靶向测序分析的合格率达到 99%。
更新日期：2024-05-24
中文翻译：
基于多个在线主动阀的离心微流体用于病毒核酸的动态固相富集和纯化
核酸即时检测（POCT）在传染病预防和控制领域以及个性化精准医疗的发展中具有重要意义。然而，传统的核酸检测方法仍然面临检测时间长、依赖大量专业设备和人员等挑战，使其不适合床旁应用。在这里，我们提出了一种用于自动核酸提取的创新主动离心微流体系统（ACMS），包括主动阀控制和磁控制模块。根据硅胶膜和锡箔之间的弹性公差差异，设计了一种片上离心穿刺阀（PV），可根据需要释放预嵌入的液体试剂。此外，我们还利用可返回阀（RV）技术，利用硅胶膜的高弹性公差来精确控制液体的保留和释放。通过采用在线可控磁阀，我们实现了磁珠的可控、快速聚集和分散。最终的芯片封装了核酸提取所需的多种试剂和磁珠。样品添加并加载到仪器中后，30分钟内即可完成芯片上的自动化样品加载和核酸提取、纯化和收集，使整体操作时间减少一半，甚至将假病毒提取效率提高三个数量级。 因此，实时荧光定量 PCR 扩增已成功检测到 SARS-CoV-2 病毒的多个靶标（检测限低至每 μL 10 个拷贝），并且靶向测序分析的合格率达到 99%。
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