A portable, rapid isothermal amplification kit enabling naked eye detection of SARS-CoV-2 RNAs

Since the coronavirus disease 2019 (COVID-19) pandemic, isothermal amplification techniques have attracted attention due to their higher sensitivity and specificity, compared with immunoassays, and their potential application for point-of-care testing (POCT). A requirement of isothermal amplification-based POCT kits is the inclusion of a heating source with an electrical power supply. We developed an amplification-based rapid kit, which is a portable and naked eye-detectable reverse transcriptase (RT)-recombinase polymerase amplification (RPA) kit. The rapid RT-RPA kit consists of a flow-controllable paper chip, a nickel-chromium (NiCr)-based Joule-heating thin film, and a small-sized portable battery. We found that the Joule-heating thin film, powered by a lithium-ion battery (7.5 g, 20 mm × 35 mm size), was able to maintain the required temperature for the RPA reaction. After the RPA reaction, which takes approximately 20 min, the flow-controllable paper chip automatically enabled visualization of the amplicon by time-delayed release of gold nanoparticle-based optical probes. Using this system, we successfully detected severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) at levels as low as 10 copies μL^-1, within 30 min.