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. We developed a molecular diagnostic test for SARS-CoV-2, FIND (Fast Isothermal Nucleic acid Detection), based on an enhanced isothermal recombinase polymerase amplification reaction. FIND has a detection limit on patient samples close to that of RT-qPCR, requires minimal instrumentation, and is highly scalable and cheap. It can be performed in high throughput, does not cross-react with other common coronaviruses, avoids bottlenecks caused by the current worldwide shortage of RNA isolation kits, and takes ~45 minutes from sample collection to results. FIND can be adapted to future novel viruses in days once sequence is available. One sentence summary Sensitive, specific, rapid, scalable, enhanced isothermal amplification method for detecting SARS-CoV-2 from patient samples.