Development of multiplexed reverse-transcription loop-mediated isothermal amplification for detection of SARS-CoV-2 and influenza viral RNA

The ongoing pandemic has demonstrated the utility of widespread surveillance and diagnostic detection of the novel SARS-CoV-2. Reverse-transcription loop-mediated isothermal amplification (RT-LAMP) has enabled broader testing, but current LAMP tests only detect single targets and require separate reactions for controls. With flu season in the Northern Hemisphere, the ability to screen for multiple targets will be increasingly important, and the ability to include internal controls in RT-LAMP allows for improved efficiency. Here we describe multiplexed RT-LAMP with four targets (SARS-CoV-2, influenza A, influenza B, human RNA) in a single reaction using real-time and end point fluorescence detection. Such increased functionality of RT-LAMP will enable even broader adoption of this molecular testing approach and aid in the fight against this public health threat. METHOD SUMMARY This study describes enhancing loop-mediated isothermal amplification through multiplexed real-time and end point detection of SARS-CoV-2 combined with influenza and control targets. By enabling multiple target detection, loop-mediated isothermal amplification can be even more widely used for diagnostics in settings where multiple viral targets are potential infectious agents and where higher-throughput testing is advantageous.

Automated summary

This study enhances loop-mediated isothermal amplification through multiplexed real-time and end point detection of SARS-CoV-2 combined with influenza and control targets. By enabling multiple target detection, loop-mediated isothermal amplification can be even more widely used for diagnostics in settings where multiple viral targets are potential infectious agents and where higher-throughput testing is advantageous.