Ding Xiong, Li Ziyue, Liu Changchun
Department of Biomedical Engineering, University of Connecticut Health Center, 263 Farmington Ave., Farmington, CT 06030, United States.
Department of Biomedical Engineering, University of Connecticut, 260 Glenbrook Road, Storrs, CT 06029, United States.
Sens Actuators B Chem. 2022 Jan 15;351:130998. doi: 10.1016/j.snb.2021.130998. Epub 2021 Oct 27.
Multiplexed detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) rather than detection targeting a single gene is crucial to ensure more accurate coronavirus disease 2019 (COVID-19) diagnostics. Here, we develop a monolithic, 3D-printed, lab-on-disc platform for multiplexed molecular detection of SARS-CoV-2. The centrifugal lab-on-disc is fabricated in one step using simple 3D printing technology, circumventing the need for aligning and binding multiple layers. By combining isothermal amplification technology, this lab-on-disc platform is capable of simultaneously detecting the nucleoprotein and envelope genes of SARS-CoV-2 as well as an internal control of the human POP7 gene. Within a 50-minute incubation period, 100 copies SARS-CoV-2 RNA can be detected through visual observation according to color and fluorescence changes in the disc. Further, we clinically validated the lab-on-disc platform by testing 20 nasopharyngeal swab samples and demonstrated a sensitivity of 100% and an accuracy of 95%. Therefore, the monolithic, 3D-printed, lab-on-disc platform provides simple, rapid, disposable, sensitive, reliable, and multiplexed molecular detection of SARS-CoV-2, holding promise for COVID-19 diagnostics at the point of care.
对严重急性呼吸综合征冠状病毒2(SARS-CoV-2)进行多重检测而非针对单个基因进行检测,对于确保更准确地诊断2019冠状病毒病(COVID-19)至关重要。在此,我们开发了一种用于SARS-CoV-2多重分子检测的整体式、3D打印的盘式实验室平台。这种离心式盘式实验室采用简单的3D打印技术一步制成,无需对齐和结合多层结构。通过结合等温扩增技术,该盘式实验室平台能够同时检测SARS-CoV-2的核蛋白基因和包膜基因以及人类POP7基因作为内部控制。在50分钟的孵育期内,通过观察盘内的颜色和荧光变化,可目视检测到100拷贝的SARS-CoV-2 RNA。此外,我们通过检测20份鼻咽拭子样本对该盘式实验室平台进行了临床验证,结果显示其灵敏度为100%,准确率为95%。因此,这种整体式、3D打印的盘式实验室平台为SARS-CoV-2提供了简单、快速、一次性、灵敏、可靠的多重分子检测,有望在护理点对COVID-19进行诊断。