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易于在集中临床实验室之外使用的快速核酸诊断。在此，我们报告了利用冻干试剂和环境温度下快速样品灭活的基于 Cas13 的核酸检测的开发和性能基准测试。我们将这些检测命名为 SHINEv.2（“简化感染突出显示以应对流行病，版本 2”），通过消除加热步骤和冷藏的需要，简化了之前报道的免 RNA 提取的 SHINEv.1 技术。试剂。SHINEv.2 使用侧流技术并在 37 ℃ 的热块中孵育，在不到 90 分钟的时间内检测到鼻咽样本中的 SARS-CoV-2，灵敏度为 90.5%，特异性为 100%（以逆转录定量聚合酶链反应为基准） °C。SHINEv.2 还允许对 Alpha、Beta、Gamma、Delta 和 Omicron SARS-CoV-2 变体进行视觉区分，并且可以通过使用体热来运行而不会造成性能损失。准确、易于使用且无需设备的核酸检测可以促进在护理点和家庭环境中对 SARS-CoV-2 和其他病原体进行更广泛的检测。