Development of Multiplexed RT-LAMP for Detection of SARS-CoV-2 and Influenza Viral RNA

Abstract

AbstractThe ongoing COVID-19 pandemic has demonstrated the utility of widespread molecular testing for surveillance and diagnostic detection of SARS-CoV-2. Reverse-transcription loop-mediated isothermal amplification (RT-LAMP) has enabled testing outside of the standard clinical laboratory PCR infrastructure, with simple and rapid tests supplementing the existing, standard methods. However, current LAMP tests have detected single targets and required separate reactions for controls or multiple targets. As flu season arrives in the Northern Hemisphere the ability to screen for multiple viral targets will be increasingly important, and the ability to include internal control assays in the RT-LAMP test allows for decreased resource use and increased throughput. Here we describe a multiplexing approach to RT-LAMP with four targets (SARS-CoV-2, Influenza A, Influenza B, and internal control human RNA) in a single reaction using real-time and endpoint fluorescence detection. This increase to the functionality of RT-LAMP will, we hope, enable even broader adoption of this power molecular testing approach to aid in the global fight against this continuing public health threat.