State Key Laboratory of Food Nutrition and Safety, Key Laboratory of Industrial Microbiology, Ministry of Education, Tianjin Key Laboratory of Industry Microbiology, National and Local United Engineering Lab of Metabolic Control Fermentation Technology, China International Science and Technology Cooperation Base of Food Nutrition/Safety and Medicinal Chemistry, College of Biotechnology, Tianjin University of Science & Technology, Tianjin, 300457, China.
State Key Laboratory of Food Nutrition and Safety, Key Laboratory of Industrial Microbiology, Ministry of Education, Tianjin Key Laboratory of Industry Microbiology, National and Local United Engineering Lab of Metabolic Control Fermentation Technology, China International Science and Technology Cooperation Base of Food Nutrition/Safety and Medicinal Chemistry, College of Biotechnology, Tianjin University of Science & Technology, Tianjin, 300457, China.
Biosens Bioelectron. 2022 Jan 1;195:113646. doi: 10.1016/j.bios.2021.113646. Epub 2021 Sep 23.
The pandemic of coronavirus disease 2019 (COVID-19) resulted from novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has become a worldwide concern. It is imperative to develop rapid, sensitive, and specific biosensing methods. Herein, we developed a CRISPR-Cas12a powered visual biosensor with a smartphone readout for ultrasensitive and selective detection of SARS-CoV-2. Simply, the SARS-CoV-2 derived nucleic acids triggered CRISPR-Cas12a based indiscriminate degradation of a single-stranded DNA that was supposed to link two gold nanoparticles, inducing the dis-aggregation of gold nanoparticles and thus generating observable color changes. This change can be readily distinguished by naked eyes as well as a smartphone with a Color Picker App. The proposed biosensor was successfully applied to detect SARS-CoV-2 gene in synthetic vectors, transcribed RNA and SARS-CoV-2 pseudoviruses. It rendered "single copy resolution" as evidenced by the 1 copy/μL limit of detection of pseudoviruses with no cross-reactivity. When the developed biosensor was challenged with SARS-CoV-2 clinical bio-samples, it provided 100% agreement (both positive and negative) with qPCR results. The sample-to-result time was roughly 90 min. Our work provides a novel and robust technology for ultrasensitive detection of SARS-CoV-2 that could be used clinically.
新型严重急性呼吸综合征冠状病毒 2（SARS-CoV-2）引起的 2019 年冠状病毒病（COVID-19）大流行已成为全球关注的焦点。开发快速、灵敏、特异的生物传感方法迫在眉睫。在此，我们开发了一种基于 CRISPR-Cas12a 的视觉生物传感器，具有智能手机读取功能，可用于超灵敏和选择性检测 SARS-CoV-2。简单地说，源自 SARS-CoV-2 的核酸触发了基于 CRISPR-Cas12a 的无差别降解，这种降解本来应该连接两个金纳米粒子，导致金纳米粒子的解聚集，从而产生可观察到的颜色变化。这种变化可以通过肉眼以及带有 Color Picker App 的智能手机轻松区分。该生物传感器成功应用于检测合成载体、转录 RNA 和 SARS-CoV-2 假病毒中的 SARS-CoV-2 基因。它实现了“单拷贝分辨率”，假病毒的检测限低至 1 拷贝/μL，无交叉反应。当用开发的生物传感器检测 SARS-CoV-2 临床生物样本时，它与 qPCR 结果完全一致（均为阳性和阴性）。从样本到结果的时间大约为 90 分钟。我们的工作为 SARS-CoV-2 的超灵敏检测提供了一种新颖而强大的技术，可用于临床。
