Rapid detection of COVID-19 using an isothermal amplification-based lateral flow assay

3月11日，由严重急性呼吸综合征冠状病毒2型(SARS-CoV-2)引起的高度传播和致命性冠状病毒疾病新冠肺炎被世界卫生组织宣布为全球大流行。建议将快速检测这种SARS-CoV-2病毒作为防止其社区传播和赢得这场抗击病毒战斗的关键。公共卫生机构广泛采用的这种病毒的金标准检测方法是基于逆转录聚合酶链式反应。虽然这种方法可靠，但它的缺点是需要复杂的仪器来进行严格的热循环，需要训练有素的人员，以及相对较长的反应时间(2-4小时)。这些都损害了加拿大每天测试大量公民的能力。为了克服这些缺点，我们提出了一种基于等温扩增机制的新型检测方法-重组酶聚合酶扩增(RPA)，它可以在恒定的低温(37~42摄氏度)下以更快的方式(&lt；20分钟)扩增核酸。与横向流动免疫分析(LFI)相结合，RPA的核酸扩增可以在几分钟内可视化，而不需要其他的扩增检测仪器。我们预计这种新的RPA-LFI方法将提供与金标准方法相当的灵敏度和特异度，同时显著减少分析时间并简化分析程序。这种新的检测方法不仅将有利于加拿大在疫情中抗击新冠肺炎，还将为在资源有限的条件下对这一公共卫生威胁进行常规监测和监测提供一种有前途的替代方案。

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.