Department of Bioengineering, Rice University, Houston, TX, USA.
Rice360 Institute for Global Health Technologies, Rice University, Houston, TX, USA.
Expert Rev Mol Diagn. 2024 Jun;24(6):509-524. doi: 10.1080/14737159.2024.2375233. Epub 2024 Jul 8.
Suitable sample collection and preparation methods are essential to enable nucleic acid amplification testing at the point of care (POC). Strategies that allow direct isothermal nucleic acid amplification testing (iNAAT) of crude sample lysate without the need for nucleic acid extraction minimize time to result as well as the need for operator expertise and costly infrastructure.
The authors review research to understand how sample matrix and preparation affect the design and performance of POC iNAATs. They focus on approaches where samples are directly combined with liquid reagents for preparation and amplification via iNAAT strategies. They review factors related to the type and method of sample collection, storage buffers, and lysis strategies. Finally, they discuss RNA targets and relevant regulatory considerations.
Limitations in sample preparation methods are a significant technical barrier preventing implementation of nucleic acid testing at the POC. The authors propose a framework for co-designing sample preparation and amplification steps for optimal performance with an extraction-free paradigm by considering a sample matrix and lytic strategy prior to an amplification assay and readout. In the next 5 years, the authors anticipate increasing priority on the co-design of sample preparation and iNAATs.
在现场即时检测（POC）中，合适的样本采集和准备方法对于核酸扩增检测至关重要。能够直接对未经核酸提取的原始样本裂解液进行等温核酸扩增检测（iNAAT）的策略，可以最大限度地减少检测结果所需的时间，减少对操作人员专业知识和昂贵基础设施的需求。
作者回顾了相关研究，以了解样本基质和制备如何影响 POC iNAAT 的设计和性能。他们重点介绍了通过 iNAAT 策略，直接将样本与液体试剂混合进行制备和扩增的方法。他们回顾了与样本采集类型和方法、储存缓冲液和裂解策略相关的因素。最后，他们讨论了 RNA 靶标和相关的监管考虑因素。
样本制备方法的局限性是阻碍核酸检测在 POC 实现的重大技术障碍。作者提出了一个框架，通过在扩增检测和读取之前，考虑样本基质和裂解策略，以无提取的范式，共同设计样本制备和扩增步骤，以实现最佳性能。在未来 5 年内，作者预计将优先考虑共同设计样本制备和 iNAAT。
