Automated sample-to-answer centrifugal microfluidic system for rapid molecular diagnostics of SARS-CoV-2
Lidija Malic, D. Brassard, Dillon Da Fonte, Christina Nassif, Maxence Mounier, André Ponton, Matthias Geißler, Matthew Shiu, Keith Morton, Teodor Veres
Abstract
, <1 h). In addition, RT-LAMP is compatible with colorimetric detection, facilitating visualization and read-out. However, even with these benefits, RT-LAMP is not yet clinically deployed at its full capacity. Lack of automation and integration of sample preparation, such as RNA extraction, limits the sensitivity and specificity of the method. Furthermore, the need for cold storage of reagents complicates its use at the point of need. The developments presented in this work address these limitations: We describe a fully automated SARS-CoV-2 detection method using RT-LAMP, which also includes up-front lysis and extraction of viral RNA, performed on a centrifugal platform with active pneumatic pumping, a disposable, all-polymer-based microfluidic cartridge and lyophilized reagents. We demonstrate that the limit of detection of the RT-LAMP assay itself is 0.2 copies per μL using N and E genes as target sequences. When combined with integrated RNA extraction, the assay sensitivity is 0.5 copies per μL, which is highly competitive to RT-qPCR. We tested the automated assay using 12 clinical swab specimens from patients and were able to distinguish positive and negative samples for SARS-CoV-2 within 60 min, thereby obtaining 100% agreement with RT-qPCR results.