Rapid detection of COVID‑19 using an isothermal amplification-based lateral flow assay
The highly transmitted and lethal coronavirus disease COVID‑19 caused by a severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been declared as a global pandemic by the World Health Organization (WHO) on March 11th. Rapid detection of this SARS-CoV-2 has been recommended as the key to prevent its community spread and to win this battle against the virus. The gold standard detection method for this virus widely adopted by public health agencies is based on reverse transcription-polymerase chain reaction. Although reliable, this method suffers the disadvantages of requiring sophisticated instrument for stringent thermal cycling, highly trained personnel, and relatively long reaction time (2-4 hours). These have impaired Canada to test a large population of citizens on a daily basis. To address these drawbacks, we proposed to develop a novel detection method based on an isothermal amplification mechanism, recombinase polymerase amplification (RPA), which can amplify nucleic acid at a constantly low temperature (37~42 degree celsius) in a much more rapid manner (<20 minutes). Combining with a lateral flow immunoassay (LFI), the nucleic acid amplicons of RPA can be visualized in a few minutes without the requirement of other instruments for amplicon detection. We anticipate this novel RPA-LFI method to provide comparable sensitivity and specificity as compared to the gold standard method, while substantially reduce the analytical time and simplify the analysis procedure. This novel detection method will not only benefit Canada to combat COVID‑19 amid the pandemic, but also provide a promising alternative for routine surveillance and monitoring of this public health threat in resource-limited conditions.
