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一种用于无实验室通用核酸提取的便携式设备的开发与验证。

Development and validation of a portable device for lab-free versatile nucleic acid extraction.

作者信息

Politza Anthony J, Liu Tianyi, Kshirsagar Aneesh, Dong Ming, Ahamed Md Ahasan, Guan Weihua

机构信息

Department of Biomedical Engineering, Pennsylvania State University, University Park, PA, USA.

Department of Electrical Engineering, Pennsylvania State University, University Park, PA, USA.

出版信息

Biotechniques. 2024 Oct;76(10):505-515. doi: 10.1080/07366205.2024.2427544. Epub 2024 Dec 2.

Abstract

Nucleic acid testing (NAT) has revolutionized diagnostics by providing precise, rapid, and scalable detection methods for diverse biological samples. These recent advancements satisfy the increasing demand for on-site diagnostics, yet sample preparation remains a significant bottleneck for achieving highly sensitive diagnostic assays. There is an unmet need for compatible, efficient, and lab-free sample preparation for point-of-care NAT. To address this, we developed a portable, lab-free, and battery-powered device for extracting nucleic acids. We explored using low centrifugal forces with existing commercial chemistry, demonstrating excellent performance. We designed and tested a battery-powered device to enable lab-free extractions, and verified reagents stored out to 6 months, suggesting exceptional deployment capabilities. We evaluated our device, comparing our results against those from a benchtop centrifuge across three types of samples: HIV RNA in buffer, HIV RNA in plasma, and SARS-CoV-2 RNA in saliva. The portable device demonstrated excellent agreement with the benchtop centrifuge, indicating high reliability. By providing an effective on-site sample preparation solution, the widespread adoption of low centrifugal extractions will improve the sensitivity and reliability of NAT and will positively impact other point-of-care technologies such as next generation sequencing (NGS), biomarker detection, and environmental monitoring.

摘要

核酸检测(NAT)通过为各种生物样本提供精确、快速且可扩展的检测方法,彻底改变了诊断方式。这些最新进展满足了对现场诊断日益增长的需求,但样本制备仍然是实现高灵敏度诊断检测的重大瓶颈。对于即时护理核酸检测,迫切需要兼容、高效且无需实验室的样本制备方法。为了解决这一问题,我们开发了一种便携式、无需实验室且由电池供电的核酸提取设备。我们探索了在现有商业化学方法中使用低离心力,结果显示性能优异。我们设计并测试了一种电池供电的设备,以实现无需实验室的提取,并验证了储存长达6个月的试剂,这表明其具有出色的部署能力。我们对我们的设备进行了评估,将我们的结果与台式离心机在三种类型样本上的结果进行了比较:缓冲液中的HIV RNA、血浆中的HIV RNA以及唾液中的SARS-CoV-2 RNA。该便携式设备与台式离心机的结果高度一致,表明其具有高可靠性。通过提供一种有效的现场样本制备解决方案,低离心力提取方法的广泛应用将提高核酸检测的灵敏度和可靠性,并将对其他即时护理技术产生积极影响,如下一代测序(NGS)、生物标志物检测和环境监测。

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