Broad Institute of Massachusetts Institute of Technology (MIT) and Harvard, Cambridge, MA, USA.
Harvard-MIT Program in Health Sciences and Technology, Cambridge, MA, USA.
Nat Biomed Eng. 2022 Aug;6(8):932-943. doi: 10.1038/s41551-022-00889-z. Epub 2022 May 30.
The widespread transmission and evolution of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) call for rapid nucleic acid diagnostics that are easy to use outside of centralized clinical laboratories. Here we report the development and performance benchmarking of Cas13-based nucleic acid assays leveraging lyophilised reagents and fast sample inactivation at ambient temperature. The assays, which we named SHINEv.2 (for 'streamlined highlighting of infections to navigate epidemics, version 2'), simplify the previously reported RNA-extraction-free SHINEv.1 technology by eliminating heating steps and the need for cold storage of the reagents. SHINEv.2 detected SARS-CoV-2 in nasopharyngeal samples with 90.5% sensitivity and 100% specificity (benchmarked against the reverse transcription quantitative polymerase chain reaction) in less than 90 min, using lateral-flow technology and incubation in a heat block at 37 °C. SHINEv.2 also allows for the visual discrimination of the Alpha, Beta, Gamma, Delta and Omicron SARS-CoV-2 variants, and can be run without performance losses by using body heat. Accurate, easy-to-use and equipment-free nucleic acid assays could facilitate wider testing for SARS-CoV-2 and other pathogens in point-of-care and at-home settings.
严重急性呼吸综合征冠状病毒 2 (SARS-CoV-2) 的广泛传播和进化需要快速易用的核酸诊断方法，以便在集中式临床实验室之外进行。在这里，我们报告了基于 Cas13 的核酸检测方法的开发和性能基准测试，该方法利用冻干试剂和在环境温度下快速样本灭活。这些检测方法被命名为 SHINEv.2（代表“简化感染检测以应对疫情，第 2 版”），通过消除加热步骤和对试剂进行冷藏的需求，简化了之前报道的无 RNA 提取的 SHINEv.1 技术。SHINEv.2 使用侧流技术和在 37°C 的热块中孵育，不到 90 分钟即可检测到鼻咽样本中的 SARS-CoV-2，灵敏度为 90.5%，特异性为 100%（与逆转录定量聚合酶链反应相比）。SHINEv.2 还允许对 Alpha、Beta、Gamma、Delta 和 Omicron SARS-CoV-2 变体进行视觉区分，并且可以在没有性能损失的情况下使用体热运行。准确、易于使用且无需设备的核酸检测方法可以促进在护理点和家庭环境中更广泛地检测 SARS-CoV-2 和其他病原体。
