Biotechnology & Bioengineering Department, Sandia National Laboratories, Livermore, CA, USA.
Expert Rev Mol Diagn. 2023 Jan;23(1):9-28. doi: 10.1080/14737159.2023.2169071. Epub 2023 Mar 1.
The SARS-CoV-2 pandemic, and the subsequent limitations on standard diagnostics, has vastly expanded the user base of Reverse Transcription Loop-mediated isothermal Amplification (RT-LAMP) in fundamental research and development. RT-LAMP has also penetrated commercial markets, with emergency use authorizations for clinical diagnosis.
This review discusses the role of RT-LAMP within the context of other technologies like RT-qPCR and rapid antigen tests, progress in sample preparation strategies to enable simplified workflow for RT-LAMP directly from clinical specimens, new challenges with primer and assay design for the evolving pandemic, prominent detection modalities including colorimetric and CRISPR-mediated methods, and translational research and commercial development of RT-LAMP for clinical applications.
RT-LAMP occupies a middle ground between RT-qPCR and rapid antigen tests. The simplicity approaches that of rapid antigen tests, making it suitable for point-of-care use, but the sensitivity nears that of RT-qPCR. RT-LAMP still lags RT-qPCR in fundamental understanding of the mechanism, and the interplay between sample preparation and assay performance. Industry is now beginning to address issues around scalability and usability, which could finally enable LAMP and RT-LAMP to find future widespread application as a diagnostic for other conditions, including other pathogens with pandemic potential.
SARS-CoV-2 大流行,以及随后对标准诊断的限制,极大地扩大了逆转录环介导等温扩增(RT-LAMP)在基础研究和开发中的用户基础。RT-LAMP 也已经渗透到商业市场,获得了临床诊断的紧急使用授权。
本文讨论了 RT-LAMP 在其他技术(如 RT-qPCR 和快速抗原检测)背景下的作用,探讨了简化 RT-LAMP 工作流程的样本制备策略的进展,使其能够直接从临床标本中进行,探讨了针对不断演变的大流行的引物和检测设计方面的新挑战,介绍了包括比色法和 CRISPR 介导方法在内的突出检测模式,以及 RT-LAMP 用于临床应用的转化研究和商业开发。
RT-LAMP 在 RT-qPCR 和快速抗原检测之间占据中间位置。其简单性类似于快速抗原检测,使其适合于即时护理用途,但敏感性接近 RT-qPCR。在对机制的基本理解方面,RT-LAMP 仍落后于 RT-qPCR,以及样品制备和分析性能之间的相互作用。现在行业开始解决可扩展性和可用性方面的问题,这最终可能使 LAMP 和 RT-LAMP 找到未来作为其他疾病(包括具有大流行潜力的其他病原体)诊断的广泛应用。