Laboratory of Molecular Diagnostics, College of Pharmaceutical Sciences, Soochow University, Suzhou 215000, Jiangsu, People's Republic of China.
Suzhou Institute for Food Control, Suzhou 215104, Jiangsu, People's Republic of China.
Bioconjug Chem. 2023 Jun 21;34(6):1147-1156. doi: 10.1021/acs.bioconjchem.3c00181. Epub 2023 May 12.
Noroviruses (NoVs) are one of the leading causes of acute gastroenteritis in humans. This study combined reverse transcription recombinase polymerase amplification (RT-RPA) with a clustered regularly interspaced short palindromic repeat/CRISPR-associated protein (CRISPR/Cas) nucleic acid detection system to develop a point-of-care testing (POCT) technology for typing NoVs. The detection can be completed within 35 min at 37 °C, covering each genotype of genogroup I (GI) and II (GII) NoVs. The sensitivity of this method is 10 copies/μL for GI and 1 copy/μL for GII NoV plasmids. For the detection of clinical samples, the detection results of this method for NoV infected samples are consistent with the RT-qPCR detection method in the laboratory, and this detection method has no cross-reactivity with rotavirus and adenovirus. Therefore, the detection method established in this study enables the diagnosis and screening of suspected patients and close contacts by POCT, which is important for the timely identification and control of NoV outbreaks. In addition, the typing detection of GI and GII NoVs can achieve a precise diagnosis and treatment of patients infected with NoVs.
诺如病毒（NoV）是导致人类急性肠胃炎的主要原因之一。本研究将逆转录重组酶聚合酶扩增（RT-RPA）与成簇规律间隔短回文重复/CRISPR 相关蛋白（CRISPR/Cas）核酸检测系统相结合，开发了一种用于诺如病毒分型的即时检测（POCT）技术。该检测可在 37°C 下 35 分钟内完成，涵盖了基因Ⅰ组（GI）和基因Ⅱ组（GII）NoV 的每个基因型。该方法的灵敏度对于 GI NoV 质粒为 10 拷贝/μL，对于 GII NoV 质粒为 1 拷贝/μL。对于临床样本的检测，该方法对感染 NoV 的样本的检测结果与实验室的 RT-qPCR 检测方法一致，并且该检测方法与轮状病毒和腺病毒无交叉反应。因此，本研究中建立的检测方法能够通过 POCT 对疑似患者和密切接触者进行诊断和筛查，这对于及时识别和控制 NoV 暴发非常重要。此外，GI 和 GII NoV 的分型检测可实现对 NoV 感染患者的精确诊断和治疗。
