Institute of Smart Farm, Gyeongsang National University, Jinju, 52828, Gyeongnam, Korea.
Division of Applied Life Science, Graduate School, Gyeongsang National University, Jinju, 52828, Gyeongnam, Korea.
Anal Bioanal Chem. 2022 Sep;414(23):6723-6733. doi: 10.1007/s00216-022-04247-5. Epub 2022 Aug 5.
Noroviruses (NoVs) are the most common causes of epidemic gastroenteritis, responsible for at least 50% of all gastroenteritis outbreaks worldwide and significant causes of foodborne illness. In the USA, approximately 21 million illnesses attributable to NoVs have annually occurred. Therefore, there is a great demand to develop a rapid, low-cost, and accurate detection method for NoVs. This study first reported colorimetric helicase-dependent amplification (HDA) methods based on specific primers integrated with HRPzyme for the rapid and sensitive detection of NoV GI and GII. The colorimetric HDA methods exhibited a detection limit of 10 copies mL of each NoV GI and GII and were confirmed to be specific to each NoV GI and GII. The period required to complete the HDA method was 2 h, including a step of RNA extraction and cDNA synthesis without expensive instruments such as a thermal cycler and detector. The cutoff value of the method for the oyster artificially inoculated with a known amount of NoV was all 10 copies g for NoV GI and GII. Therefore, the HDA method developed in this study can be useful tool for the on-site detection of NoVs in food samples.
诺如病毒（NoV）是引起流行性肠胃炎的最常见原因，至少造成了全球 50%的肠胃炎爆发，也是食源性疾病的重要致病源。在美国，每年因 NoV 而发病的人数约为 2100 万。因此，人们对开发一种快速、低成本、准确的 NoV 检测方法有着巨大的需求。本研究首次报道了基于与 HRPzyme 整合的特异性引物的比色解旋酶依赖性扩增（HDA）方法，用于快速灵敏地检测 NoV GI 和 GII。比色 HDA 方法对每个 NoV GI 和 GII 的检测限均为 10 拷贝 mL，并被证实对每个 NoV GI 和 GII 均具有特异性。完成 HDA 方法所需的时间为 2 小时，包括 RNA 提取和 cDNA 合成步骤，无需热循环仪和检测器等昂贵仪器。该方法对人工接种已知量 NoV 的牡蛎的检测截止值均为 NoV GI 和 GII 的 10 拷贝 g。因此，本研究中开发的 HDA 方法可作为现场检测食品样本中 NoV 的有用工具。
