Point-of-care testing of “sample in, answer out” is urgently needed for communicable diseases. Recently, rapid nucleic acid tests for infectious diseases have been developed for use in resource-limited areas, but they require types of equipment in central laboratories and are poorly integrated. In this work, a portable centrifugal microfluidic testing system is developed, integrated with magnetic bead-based nucleic acid extraction, recombinase-assisted amplification and CRISPR–Cas13a detection. The system, with the advantage of its power-supplied active rotating chip and highly programable flow control through integrated addressable active thermally-triggered wax valves, has a rapid turnaround time within 45 min, requiring only one user step. All reagents are preloaded into the chip and can be automatically released. By exploiting a multichannel chip, it is capable of simultaneously detecting 10 infectious viruses with limits of detection of 1 copy per reaction and 5 copies per reaction in plasmid samples and mock plasma samples, respectively. The system was used to analyse clinical plasma samples with good consistency compared to laboratory-based molecular testing. Moreover, the generalizability of our device is reported by successfully testing nasopharyngeal swabs and whole blood samples. The portable device does not require the operation of professional technicians, making it an excellent assay for on-site testing.

中文翻译：

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用于基于 CRISPR-Cas13a 的完全集成多重核酸检测的便携式一体化微流控系统


传染病迫切需要“采样、回答”的即时检测。最近，针对传染病的快速核酸检测已被开发用于资源有限的地区，但它们需要中心实验室的设备类型并且集成度差。在这项工作中，开发了一种便携式离心微流控测试系统，集成了基于磁珠的核酸提取、重组酶辅助扩增和CRISPR-Cas13a检测。该系统凭借其供电主动旋转芯片和通过集成可寻址主动热触发蜡阀进行高度可编程流量控制的优势，可在 45 分钟内快速完成周转，只需用户执行一个步骤。所有试剂均预装至芯片内并可自动释放。通过利用多通道芯片，它能够同时检测 10 种传染性病毒，质粒样本和模拟血浆样本中的检测限分别为每个反应 1 个拷贝和每个反应 5 个拷贝。与基于实验室的分子检测相比，该系统用于分析临床血浆样本，具有良好的一致性。此外，通过成功测试鼻咽拭子和全血样本，报告了我们的设备的普遍性。该便携式设备不需要专业技术人员操作，非常适合现场测试。