College of Food Science and Technology, Shanghai Ocean University, Shanghai, China.
Laboratory for Marine Biology and Biotechnology, Qingdao National Laboratory for Marine Science and Technology, Qingdao, China.
Arch Virol. 2020 Dec;165(12):2767-2776. doi: 10.1007/s00705-020-04798-x. Epub 2020 Sep 19.
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.
人类诺如病毒是全球病毒性肠胃炎的主要病因。快速检测有助于疾病爆发的管理，但由于缺乏可靠和便携的方法，现场诊断较为困难。重组酶聚合酶扩增（RPA）是一种强大的等温扩增方法，能够使用简单的设备快速扩增和检测核酸。在这项研究中，建立并评估了与人类基因 II 组（GII）诺如病毒特异性侧向流动（LF）条相结合的 RPA。该检测方法可特异性检测纯化的 GII 诺如病毒以及煮沸的人类粪便样本中的 RNA，反应的灵敏度为每反应 50 个诺如病毒基因组拷贝。一步 RT-RPA-LF 的整个检测过程在 20 分钟内完成，比标准实时 RT-PCR 快八倍。这里描述的 RT-RPA-LF 方法适用于快速现场诊断人类粪便样本中的所有 GII 诺如病毒。
