Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA, USA.
Institute for Medical Engineering and Science and Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA.
Nat Biomed Eng. 2022 Aug;6(8):968-978. doi: 10.1038/s41551-022-00919-w. Epub 2022 Aug 8.
Rapid, accurate and frequent detection of the RNA of SARS-CoV-2 (severe acute respiratory syndrome coronavirus 2) and of serological host antibodies to the virus would facilitate the determination of the immune status of individuals who have Coronavirus disease 2019 (COVID-19), were previously infected by the virus, or were vaccinated against the disease. Here we describe the development and application of a 3D-printed lab-on-a-chip that concurrently detects, via multiplexed electrochemical outputs and within 2 h, SARS-CoV-2 RNA in saliva as well as anti-SARS-CoV-2 immunoglobulins in saliva spiked with blood plasma. The device automatedly extracts, concentrates and amplifies SARS-CoV-2 RNA from unprocessed saliva, and integrates the Cas12a-based enzymatic detection of SARS-CoV-2 RNA via isothermal nucleic acid amplification with a sandwich-based enzyme-linked immunosorbent assay on electrodes functionalized with the Spike S1, nucleocapsid and receptor-binding-domain antigens of SARS-CoV-2. Inexpensive microfluidic electrochemical sensors for performing multiplexed diagnostics at the point of care may facilitate the widespread monitoring of COVID-19 infection and immunity.
快速、准确、频繁地检测 SARS-CoV-2（严重急性呼吸系统综合征冠状病毒 2）的 RNA 和血清宿主对该病毒的抗体，将有助于确定患有 2019 年冠状病毒病（COVID-19）、先前感染过该病毒或接种过该疾病疫苗的个体的免疫状态。在这里，我们描述了一种 3D 打印的芯片实验室的开发和应用，该实验室通过多路电化学输出，在 2 小时内同时检测唾液中的 SARS-CoV-2 RNA 以及含有血浆的唾液中的抗 SARS-CoV-2 免疫球蛋白。该设备可自动从未经处理的唾液中提取、浓缩和扩增 SARS-CoV-2 RNA，并将基于 Cas12a 的 SARS-CoV-2 RNA 酶促检测与基于电极的夹心酶联免疫吸附试验相结合，该电极功能化有 SARS-CoV-2 的 Spike S1、核衣壳和受体结合域抗原，用于进行等温核酸扩增。用于在护理点进行多重诊断的廉价微流体电化学传感器可能有助于广泛监测 COVID-19 感染和免疫。
