An enhanced isothermal amplification assay for viral detection

Qian, Jason; Boswell, Sarah A.; Chidley, Christopher; Lu, Zhi-xiang; Pettit, Mary E.; Gaudio, Benjamin L.; Fajnzylber, Jesse M.; Ingram, Ryan T.; Ward, Rebecca H.; Li, Jonathan Z.; Springer, Michael

Rapid, inexpensive, robust diagnostics are essential to control the spread of infectious diseases. Current state of the art diagnostics are highly sensitive and specific, but slow, and require expensive equipment. Here we report the development of a molecular diagnostic test for SARS-CoV-2 based on an enhanced recombinase polymerase amplification (eRPA) reaction. eRPA has a detection limit on patient samples down to 5 viral copies, requires minimal instrumentation, and is highly scalable and inexpensive. eRPA does not cross-react with other common coronaviruses, does not require RNA purification, and takes ~45 min from sample collection to results. eRPA represents a first step toward at-home SARS-CoV-2 detection and can be adapted to future viruses within days of genomic sequence availability.