Low-Cost, Open-Source Device for High-Performance Fluorescence Detection of Isothermal Nucleic Acid Amplification Reactions,ACS Biomaterials Science & Engineering

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Low-Cost, Open-Source Device for High-Performance Fluorescence Detection of Isothermal Nucleic Acid Amplification Reactions
ACS Biomaterials Science & Engineering ( IF 5.4 ) Pub Date : 2021-09-14 , DOI: 10.1021/acsbiomaterials.1c01105
Andrew H Buultjens ₁ , Koen Vandelannoote ₁ , Liam K Sharkey ₁ , Benjamin P Howden _{2,

3,

4} , Ian R Monk ₁ , Jean Y H Lee _{1,

5} , Timothy P Stinear _{1,

3}

Affiliation

The ability to detect SARS-CoV-2 is critical to implementing evidence-based strategies to address the COVID-19 global pandemic. Expanding SARS-CoV-2 diagnostic ability beyond well-equipped laboratories widens the opportunity for surveillance and control efforts. However, such advances are predicated on the availability of rapid, scalable, accessible, yet high-performance diagnostic platforms. Methods to detect viral RNA using reverse transcription loop-mediated isothermal amplification (RT-LAMP) show promise as rapid and field-deployable tests; however, the per-unit costs of the required diagnostic hardware can be a barrier for scaled deployment. Here, we describe a diagnostic hardware configuration for LAMP technology, named the FABL-8, that can be built for approximately US$380 per machine and provide results in under 30 min. Benchmarking showed that FABL-8 has a similar performance to a high-end commercial instrument for detecting fluorescence-based LAMP reactions. Performance testing of the instrument with RNA extracted from a SARS-CoV-2 virus dilution series revealed an analytical detection sensitivity of 50 virus copies per microliter—a detection threshold suitable to detect patient viral load in the first few days following symptom onset. In addition to the detection of SARS-CoV-2, we show that the system can be used to detect the presence of two bacterial pathogens, demonstrating the versatility of the platform for the detection of other pathogens. This cost-effective and scalable hardware alternative allows democratization of the instrumentation required for high-performance molecular diagnostics, such that it could be available to laboratories anywhere—supporting infectious diseases surveillance and research activities in resource-limited settings.

中文翻译：

用于等温核酸扩增反应的高性能荧光检测的低成本、开源设备

检测 SARS-CoV-2 的能力对于实施基于证据的策略来应对 COVID-19 全球大流行至关重要。将 SARS-CoV-2 诊断能力扩展到设备齐全的实验室之外，扩大了监测和控制工作的机会。然而，这些进步取决于快速、可扩展、可访问且高性能的诊断平台的可用性。使用逆转录环介导的等温扩增 (RT-LAMP) 检测病毒 RNA 的方法显示出作为快速和可现场部署的测试的前景；然而，所需诊断硬件的单位成本可能成为大规模部署的障碍。在这里，我们描述了 LAMP 技术的诊断硬件配置，名为FABL-8，每台机器的成本约为 380 美元，并在 30 分钟内提供结果。基准测试表明FABL-8具有与用于检测基于荧光的 LAMP 反应的高端商业仪器相似的性能。使用从 SARS-CoV-2 病毒稀释系列中提取的 RNA 对该仪器的性能测试显示，分析检测灵敏度为每微升 50 个病毒拷贝——该检测阈值适用于在症状出现后的最初几天内检测患者病毒载量。除了检测 SARS-CoV-2 之外，我们还展示了该系统可用于检测两种细菌病原体的存在，证明了该平台在检测其他病原体方面的多功能性。这种具有成本效益且可扩展的硬件替代方案允许高性能分子诊断所需的仪器大众化，

更新日期：2021-10-12

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