London Biofoundry, Imperial College Translation and Innovation Hub, White City Campus, 80 Wood Lane, London, W12 0BZ, UK.
Section of Structural and Synthetic Biology, Department of Infectious Disease, Imperial College London, London, SW7 2AZ, UK.
Nat Commun. 2020 Sep 8;11(1):4464. doi: 10.1038/s41467-020-18130-3.
The SARS-CoV-2 pandemic has shown how a rapid rise in demand for patient and community sample testing can quickly overwhelm testing capability globally. With most diagnostic infrastructure dependent on specialized instruments, their exclusive reagent supplies quickly become bottlenecks, creating an urgent need for approaches to boost testing capacity. We address this challenge by refocusing the London Biofoundry onto the development of alternative testing pipelines. Here, we present a reagent-agnostic automated SARS-CoV-2 testing platform that can be quickly deployed and scaled. Using an in-house-generated, open-source, MS2-virus-like particle (VLP) SARS-CoV-2 standard, we validate RNA extraction and RT-qPCR workflows as well as two detection assays based on CRISPR-Cas13a and RT-loop-mediated isothermal amplification (RT-LAMP). In collaboration with an NHS diagnostic testing lab, we report the performance of the overall workflow and detection of SARS-CoV-2 in patient samples using RT-qPCR, CRISPR-Cas13a, and RT-LAMP. The validated RNA extraction and RT-qPCR platform has been installed in NHS diagnostic labs, increasing testing capacity by 1000 samples per day.
SARS-CoV-2 大流行表明，患者和社区样本检测需求的快速增长如何迅速使全球检测能力不堪重负。由于大多数诊断基础设施依赖于专用仪器，其独家试剂供应很快成为瓶颈，因此迫切需要采用方法来提高检测能力。我们通过将伦敦生物制造厂重新聚焦于替代检测管道的开发来应对这一挑战。在这里，我们提出了一种试剂不可知的自动化 SARS-CoV-2 检测平台，该平台可以快速部署和扩展。我们使用内部生成的开源 MS2 病毒样颗粒 (VLP) SARS-CoV-2 标准，验证了 RNA 提取和 RT-qPCR 工作流程以及基于 CRISPR-Cas13a 和 RT 环介导等温扩增 (RT-LAMP) 的两种检测方法。我们与 NHS 诊断检测实验室合作，报告了使用 RT-qPCR、CRISPR-Cas13a 和 RT-LAMP 对患者样本进行整个工作流程和 SARS-CoV-2 检测的性能。经过验证的 RNA 提取和 RT-qPCR 平台已安装在 NHS 诊断实验室中，每天可增加 1000 个样本的检测能力。
