College of Biomass Science and Engineering, Department of Respiratory and Critical Care Medicine, State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, Sichuan University, Chengdu, China.
Department of Chemistry, Key Laboratory of Bioorganic Phosphorus Chemistry and Chemical Biology, Tsinghua University, Beijing, China.
Nat Biomed Eng. 2022 Aug;6(8):957-967. doi: 10.1038/s41551-022-00907-0. Epub 2022 Jul 14.
The evolution of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has highlighted the need for versatile diagnostic assays that can discriminate among emerging variants of the virus. Here we report the development and performance benchmarking of an inexpensive (approximately US$0.30 per test) assay for the rapid (sample-to-answer time within 30 min) colorimetric detection of SARS-CoV-2 variants. The assay, which we integrated into foldable paper strips, leverages nucleic acid strand-displacement reactions, the thermodynamic energy penalty associated with single-base-pair mismatches and the metal-ion-controlled enzymatic cleavage of urea to amplify the recognition of viral RNAs for the colorimetric readout of changes in pH via a smartphone. For 50 throat swab samples, the assay simultaneously detected the presence of SARS-CoV-2 and mutations specific to the SARS-CoV-2 variants Alpha, Beta and Gamma, with 100% concordance with real-time quantitative polymerase chain reaction and RNA sequencing. Customizable and inexpensive paper-based assays for the detection of viruses and their variants may facilitate viral surveillance.
严重急性呼吸综合征冠状病毒 2 (SARS-CoV-2) 的进化突显出需要多功能的诊断检测方法，以区分病毒的新兴变体。在这里，我们报告了一种廉价（每个测试约 0.30 美元）的检测方法的开发和性能基准测试，用于快速（30 分钟内完成样本到答案的时间）比色检测 SARS-CoV-2 变体。该检测方法我们集成到可折叠的纸带上，利用核酸链置换反应、与单碱基对不匹配相关的热力学能量惩罚以及金属离子控制的脲酶切割，通过智能手机放大对病毒 RNA 的识别，从而对 pH 值变化进行比色读出。对于 50 个咽喉拭子样本，该检测方法同时检测到 SARS-CoV-2 的存在和 SARS-CoV-2 变体 Alpha、Beta 和 Gamma 的特定突变，与实时定量聚合酶链反应和 RNA 测序的一致性为 100%。用于检测病毒及其变体的定制和廉价的基于纸张的检测方法可能有助于病毒监测。
