A microfluidic system for rapid nucleic acid analysis based on real-time convective PCR at point-of-care testing.
A microfluidic system for rapid nucleic acid analysis based on real-time convective PCR at point-of-care testing.
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一种基于实时对流 PCR 的即时检测快速核酸分析微流体系统。
DOI：
10.1007/s10404-022-02577-5
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发表时间：
2022
影响因子：
2.8
通讯作者：
中科院分区：
文献类型：
--
作者：
建立了一种基于实时对流PCR的核酸快速分析微流控系统。为了实现“进样、出样”的核酸分析，开发了一种微流控芯片，用于高效提取核酸，同时应用对流PCR（CPCR）进行快速核酸扩增。利用由试剂预储存室、裂解和洗涤室、洗脱室和废物室组成的集成微流控芯片，可以在有限的时间内对大尺寸的测试样品自动进行基于磁珠的核酸提取。基于易于操作的策略，可以方便地释放不同的预存储试剂，以用于不同步骤的连续反应。为了实现有效的混合，开发了一种便携式配套装置，以在核酸提取中将适当控制的3-D致动引入磁珠。在CPCR扩增中，PCR试剂可以在反应器的热区和冷区之间自发地和重复地循环，用于基于伪等温加热的空间域热循环。开发了一种手持式实时CPCR装置，用于进行核酸扩增和原位检测。为了扩展检测通量，多个手持式实时CPCR设备可以通过公共控制系统分组在一起。结果表明，该微流控系统能够成功检测到浓度低至1.0 TCID 50/ml的甲型H1N1流感病毒。
A microfluidic system for rapid nucleic acid analysis based on real-time convective PCR is developed. To perform ‘sample-in, answer-out’ nucleic acid analysis, a microfluidic chip is developed to efficiently extract nucleic acid, and meanwhile convective PCR (CPCR) is applied for rapid nucleic acid amplification. With an integrated microfluidic chip consisting of reagent pre-storage chambers, a lysis & wash chamber, an elution chamber and a waste chamber, nucleic acid extraction based on magnetic beads can be automatically performed for a large size of test sample within a limited time. Based on an easy-to-operate strategy, different pre-stored reagents can be conveniently released for consecutive reaction at different steps. To achieve efficient mixing, a portable companion device is developed to introduce properly controlled 3-D actuation to magnetic beads in nucleic acid extraction. In CPCR amplification, PCR reagent can be spontaneously and repeatedly circulated between hot and cool zones of the reactor for space-domain thermal cycling based on pseudo-isothermal heating. A handheld real-time CPCR device is developed to perform nucleic acid amplification and in-situ detection. To extend the detection throughput, multiple handheld real-time CPCR devices can be grouped together by a common control system. It is demonstrated that influenza A (H1N1) viruses with the reasonable concentration down to 1.0 TCID50/ml can be successfully detected with the microfluidic system.
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