Development of a paper-based POCT device for RT-RPA of SARS-CoV-2

Abstract

As the SARS-CoV-2 spreads as a pandemic, the importance of fast and accurate diagnosis is being emphasized. Several biosensors have appeared for rapid and accurate target detection, among which PCR is the most common COVID-19 diagnostics test today. However, there is a disadvantage that point of care testing(POCT) is impossible because expensive equipment and trained technician are required. To overcome these issue, the isothermal amplification method has emerged. Among the isothermal amplification methods, recombinase polymerase amplification (RPA) has a low required temperature (37℃~42℃) and rapid amplification (20 minutes), so RPA is a suitable test method as a POCT. We developed molecular diagnostics that combines RT-RPA with lateral flow assay (LFA), which is detectable in color with a low voltage within 30 minutes. The device we developed succeeded in detecting 103 copy µL-1 of purified SARS-CoV-2 and the simple primer design is expected to be a powerful tool that can quickly respond to outbreaks of other infectious diseases.