Sample-to-Answer Microfluidic Nucleic Acid Testing (NAT) on Lab-on-a-Disc for Malaria Detection at Point of Need.
Sample-to-Answer Microfluidic Nucleic Acid Testing (NAT) on Lab-on-a-Disc for Malaria Detection at Point of Need.
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DOI：
10.1007/978-1-0716-1803-5_16
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发表时间：
2022
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对于现场部署的NAT来说，最大的挑战之一与检测的前端有关--从原始样本中提取核酸。理想的核酸样品制备应该是简单、可伸缩和易于操作的。在本章中，我们将介绍一种用于样本到答案疟疾诊断的光盘实验室NAT设备。寄生虫DNA样本制备和随后的实时LAMP检测无缝集成在一张一次性单个微流控光盘上，由节能的非离心机磁场相互作用驱动。每个光盘包含四个平行的测试单元，可以配置为四个相同的测试或四个特定物种的测试。当配置为物种特异性测试时，它可以识别两个最具生命威胁的疟疾物种(恶性疟原虫和间日疟原虫)。带有4路分析仪的试剂盘(在第1章中讨论)能够在40分钟的周转时间内同时处理4个样品。该方法对L全血的检出限为0.5个寄生虫/μ，足以检测出无症状的寄生虫携带者。化验是使用配套章节中描述的自动化设备进行的。灵敏度、特异度、成本和可扩展的样品制备相结合，表明实时荧光灯设备可能对现场疟疾筛查特别有用。
One of the grand challenges for field-deployable NATs is related to the front end of the assays – nucleic acid extraction from raw samples. The ideal nucleic acid sample preparation should be simple, scalable, and easy-to-operate. In this chapter, we present a lab-on-a-disc NAT device for sample-to-answer malaria diagnosis. The parasite DNA sample preparation and subsequent real-time LAMP detection were seamlessly integrated on a disposable single microfluidic compact disc, driven by energy-efficient, non-centrifuge based magnetic field interactions. Each disc contains four parallel testing units, which could be configured either as four identical tests or as four species-specific tests. When configured as species-specific tests, it could identify two of the most life-threatening malaria species (P. falciparum and P. vivax). The reagent disc with a 4-plex analyzer (discussed in Chapter 1) is capable of processing four samples simultaneously with 40 min turnaround time. It achieves a detection limit of ~0.5 parasites/μl for whole blood, sufficient for detecting asymptomatic parasite carriers. The assay is performed with an automated device described in the companion chapter. The combination of sensitivity, specificity, cost, and scalable sample preparation suggests the real-time fluorescence LAMP device could be particularly useful for malaria screening in the field settings.