A point-of-care microfluidic biosensing system for rapid and ultrasensitive nucleic acid detection from clinical samples
A point-of-care microfluidic biosensing system for rapid and ultrasensitive nucleic acid detection from clinical samples
复制标题
用于从临床样本中快速、超灵敏地检测核酸的护理点微流控生物传感系统
DOI：
10.1039/d3lc00372h
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发表时间：
2023
期刊：
影响因子：
6.1
通讯作者：
Zhang, Yi
中科院分区：
文献类型：
--
作者：
Zhang, Yuxuan;Song, Yang;Weng, Zhengyan;Yang, Jie;Avery, Lori;Dieckhaus, Kevin D.;Lai, Rebecca Y.;Gao, Xue;Zhang, Yi
快速和超灵敏的即时RNA检测在各种传染病的诊断和管理中起着关键作用。逆转录-定量聚合酶链反应（RT-qPCR）的金标准检测方法非常灵敏和准确，但受到周转时间较长（1-2天）的限制。另一方面，抗原测试提供快速的家庭检测（通常约15分钟），但灵敏度低，假阴性率高。理想的即时诊断设备将联合收割机PCR水平灵敏度和与抗原检测相当的快速样品到结果工作流程的优点。然而，现有的检测平台通常具有上级灵敏度或快速的样品到结果时间，但不能同时具有两者。本文报道了一种即时微流控装置，该装置提供了对临床样品中病毒RNA的超灵敏但快速的检测。该装置包括一个用于精确操作小体积样品的微流控芯片，一个用于病毒裂解和核糖核酸酶灭活的小型化加热器，一个用于无靶向预扩增和超灵敏RNA检测的Cas 13 a电化学传感器，以及一个用于数据采集的智能手机兼容恒电位仪。作为示范，该设备实现了热灭活SARS-CoV-2样本的检测，检测限在25分钟内降至10 aM，这与RT-PCR的灵敏度和抗原检测的快速性相当。该平台还在25分钟的样本到结果时间内成功区分了所有9个阳性的未经处理的临床SARS-CoV-2鼻咽拭子样本和4个阴性样本。总之，该设备提供了一种即时护理解决方案，可以部署在实验室环境之外的各种环境中，以快速准确地检测临床样本中的RNA。该设备可以扩展到检测其他病毒目标，如人类免疫缺陷病毒自我检测和寨卡病毒，其中需要快速和超灵敏的即时检测。
Rapid and ultrasensitive point-of-care RNA detection plays a critical role in the diagnosis and management of various infectious diseases. The gold-standard detection method of reverse transcription-quantitative polymerase chain reaction (RT-qPCR) is ultrasensitive and accurate yet limited by the lengthy turnaround time (1–2 days). On the other hand, an antigen test offers rapid at-home detection (typically ~15 min) but suffers from low sensitivity and high false-negative rates. An ideal point-of-care diagnostic device would combine the merits of PCR-level sensitivity and rapid sample-to-result workflow comparable to antigen testing. However, the existing detection platforms typically possess superior sensitivity or rapid sample-to-result time, but not both. This paper reports a point-of-care microfluidic device that offers ultrasensitive yet rapid detection of viral RNA from clinical samples. The device consists of a microfluidic chip for precisely manipulating small volumes of samples, a miniaturized heater for viral lysis and ribonuclease inactivation, a Cas13a-electrochemical sensor for target preamplification-free and ultrasensitive RNA detection, and a smartphone-compatible potentiostat for data acquisition. As demonstrations, the devices achieve the detection of heat-inactivated SARS-CoV-2 samples with a limit of detection down to 10 aM within 25 minutes, which is comparable to the sensitivity of RT-PCR and rapidness of an antigen test. The platform also successfully distinguishes all nine positive unprocessed clinical SARS-CoV-2 nasopharyngeal swab samples from four negative samples within 25 minutes of sample-to-result time. Together, this device provides a point-of-care solution that can be deployed in diverse settings beyond laboratory environments for rapid and accurate detection of RNA from clinical samples. The device can potentially be expandable to detect other viral targets, such as human immunodeficiency virus self-testing and Zika virus, where rapid and ultrasensitive point-of-care detection is required.