van Beuningen Rinie, Jim Kin Ki, Boot Maikel, Ossendrijver Michel, Keijser Bart J F, van de Bovenkamp Jeroen H B, Melchers Willem J G, Kievits Tim
Stichting Therapie op Maat (TOMi Foundation), 's-Hertogenbosch, 5223DE, The Netherlands.
Department of Fundamental Microbiology, Faculty of Biology and Medicine, University of Lausanne, Lausanne, CH-1015, Switzerland.
Biol Methods Protoc. 2024 Nov 27;9(1):bpae090. doi: 10.1093/biomethods/bpae090. eCollection 2024.
The coronavirus disease 2019 (COVID-19) pandemic underscored the necessity for rapid and efficient diagnostic testing to mitigate outbreaks and control disease transmission. While real-time reverse transcriptase quantitative PCR (RT-qPCR) has been the gold standard due to its high sensitivity and specificity, its logistical complexities and extended turnaround times highlighted the need for alternative molecular methods and non-standard equipment and consumables not subject to supply chain pressure. Loop-mediated isothermal amplification (LAMP) offers several advantages over RT-qPCR, including faster processing time, assay flexibility and cost-effectiveness. During the pandemic, LAMP was successfully demonstrated as a viable alternative to RT-qPCR for SARS-Related Coronavirus 2 detection. However, due to a 100 to 1,000-fold increase in testing volumes, there was an imminent need for automating and scaling up existing LAMP testing workflows leveraging a robotic infrastructure, while retaining analytical performance and cost-effectiveness. In 2020, the Foundation TOMi started the "TOMi corona initiative" to develop and validate a high-throughput, end-to-end, automated, scalable single-step RNA purification, and LAMP-based COVID-19 testing system called SMART-LAMP (Scalable Molecular Automation for Rapid Testing using LAMP) that can process up to 40,000 samples per day using existing laboratory equipment infrastructure with sensitivity comparable to RT-qPCR. This system provides a rapid and scalable diagnostic solution for future pandemics, capable of processing over 40,000 samples per day. In addition, the system is designed to minimize consumable costs and reduces the overall use of plastics to align with increasingly strict sustainability goals that will be imposed over the coming years. Importantly, this system and public-private partnerships in the TOMi corona initiative has the potential to serve as a baseline to enhance pandemic preparedness and response capabilities.
2019年冠状病毒病（COVID-19）大流行凸显了快速高效诊断检测对于减轻疫情爆发和控制疾病传播的必要性。虽然实时逆转录定量聚合酶链反应（RT-qPCR）因其高灵敏度和特异性一直是金标准，但其后勤复杂性和较长的周转时间凸显了对不受供应链压力影响的替代分子方法以及非标准设备和耗材的需求。环介导等温扩增（LAMP）相对于RT-qPCR具有多个优势，包括处理时间更快、检测灵活性和成本效益。在大流行期间，LAMP已成功证明是用于检测严重急性呼吸综合征冠状病毒2的RT-qPCR的可行替代方法。然而，由于检测量增加了100至1000倍，迫切需要利用机器人基础设施实现现有LAMP检测工作流程的自动化和扩大规模，同时保持分析性能和成本效益。2020年，TOMi基金会启动了“TOMi冠状病毒计划”，以开发和验证一种名为SMART-LAMP（使用LAMP进行快速检测的可扩展分子自动化）的高通量、端到端、自动化、可扩展的单步RNA纯化和基于LAMP的COVID-19检测系统，该系统使用现有的实验室设备基础设施每天可处理多达40000个样本，灵敏度与RT-qPCR相当。该系统为未来的大流行提供了一种快速且可扩展的诊断解决方案，每天能够处理超过40000个样本。此外，该系统旨在将耗材成本降至最低，并减少塑料的总体使用量，以符合未来几年将实施的日益严格的可持续发展目标。重要的是，该系统以及TOMi冠状病毒计划中的公私合作伙伴关系有可能作为加强大流行防范和应对能力的基线。
