Saint-Petersburg Pasteur Institute, 197101 Saint-Petersburg, Russia.
Faculty of Molecular Biology, Moscow State University M.V. Lomonosov, 188512 Moscow, Russia.
Biosensors (Basel). 2023 Feb 10;13(2):252. doi: 10.3390/bios13020252.
Nipah virus (NiV) is a zoonotic RNA virus which infects humans and animals in Asian countries. Infection in humans occurs in different forms, from asymptomatic infection to fatal encephalitis, and death occurred in 40-70% of those infected in outbreaks that occurred between 1998 and 2018. Modern diagnostics is carried out by real-time PCR to identify pathogens or by ELISA to detect antibodies. Both technologies are labor-intensive and require the use of expensive stationary equipment. Thus, there is a need to develop alternative simple, fast and accurate test systems for virus detection. The aim of this study was to develop a highly specific and easily standardized system for the detection of Nipah virus RNA. In our work, we have developed a design for a Dz_NiV biosensor based on a split catalytic core of deoxyribozyme 10-23. It was shown that the assembly of active 10-23 DNAzymes occurred only in the presence of synthetic target Nipah virus RNA and that this was accompanied by stable fluorescence signals from the cleaved fluorescent substrates. This process was realized at 37 °C, pH 7.5, and in the presence of magnesium ions, with a 10 nM limit of detection achieved for the synthetic target RNA. Constructed via a simple and easily modifiable process, our biosensor may be used for the detection of other RNA viruses.
果蝠尼帕病毒（NiV）是一种感染亚洲国家人类和动物的动物源性 RNA 病毒。人类感染的形式不同，从无症状感染到致命脑炎，1998 年至 2018 年爆发的感染中，死亡率为 40-70%。现代诊断通过实时 PCR 识别病原体或通过 ELISA 检测抗体进行。这两种技术都需要耗费大量的人力，并且需要使用昂贵的固定设备。因此，需要开发替代的简单、快速和准确的病毒检测系统。本研究旨在开发一种高度特异性和易于标准化的尼帕病毒 RNA 检测系统。在我们的工作中，我们基于脱氧核酶 10-23 的分裂催化核心，开发了一种 Dz_NiV 生物传感器的设计。结果表明，只有在存在合成靶标尼帕病毒 RNA 的情况下，才会发生活性 10-23 DNAzyme 的组装，并且伴随着来自切割荧光底物的稳定荧光信号。该过程在 37°C、pH7.5 和镁离子存在下进行，对于合成靶 RNA 的检测限为 10 nM。通过简单且易于修饰的过程构建的生物传感器，可用于检测其他 RNA 病毒。
