Human norovirus is the leading cause of viral gastroenteritis worldwide. Rapid detection facilitates management of disease outbreaks, but field diagnosis is difficult to achieve due to the lack of reliable and portable methods. Recombinase polymerase amplification (RPA) is a robust isothermal amplification method that is capable of rapidly amplifying and detecting nucleic acids using simple equipment. In this study, RPA combined with lateral flow (LF) strips specific for human genogroup II (GII) noroviruses was established and evaluated. The assay specifically detects purified GII noroviruses as well as RNA in boiled human stool samples, with a sensitivity of 50 norovirus genome copies per reaction. The whole detection procedure of the one-step RT-RPA-LF is completed within 20 min, which is eight times faster than that of the standard real-time RT-PCR. The RT-RPA-LF method described here is suitable for rapid field diagnosis of all GII noroviruses in human stool samples.

人类诺如病毒是全世界病毒性肠胃炎的主要原因。快速检测有助于疾病爆发的管理，但是由于缺乏可靠且可移植的方法，很难实现现场诊断。重组酶聚合酶扩增（RPA）是一种可靠的等温扩增方法，能够使用简单的设备快速扩增和检测核酸。在这项研究中，建立了RPA并结合了人类基因组II（GII）诺如病毒特有的侧流（LF）条。该测定法特异性检测煮沸的人粪便样品中的纯化GII诺如病毒以及RNA，每个反应的灵敏度为50诺如病毒基因组拷贝。一步式RT-RPA-LF的整个检测过程可在20分钟内完成，比标准实时RT-PCR快八倍。本文所述的RT-RPA-LF方法适用于人粪便样本中所有GII诺如病毒的快速现场诊断。