A Simple Poc Device for Temperature Control of Multiple Reactions During Recombinase Polymerase Amplification

Aubrey Schultz, Shannon Servoss, Robert Beitle
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Abstract

For individuals with infectious diseases, early and accurate diagnosis is critical. A rapid diagnosis allows for prompt and effective treatment and increases the chance of a full recovery without complications. Additionally, when containing a wide-scale infectious disease outbreak, circumstances are significantly improved by the ability to test the populace frequently, swiftly, and affordably. Regarding specificity and sensitivity, nucleic acid amplification tests (NAAT) are one of the best options for diagnosing infectious diseases. Historically, polymerase chain reaction (PCR) has been used, but complex thermocycling and complicated PCR protocols have often limited PCR to clinical settings. Due to increased simplicity, the isothermal NAAT recombinase polymerase amplification (RPA) has the potential to deliver reliable POC diagnostics in low-resource settings. When designing POC devices for isothermal NAATs, creating isothermal temperature conditions is perhaps the most significant challenge. This work presents a flexible and robust device capable of incubating 3 RPA reactions for simultaneous amplification in conditions conducive to POC testing. The device costs ~$ 60 USD to construct and is easy to assemble. A battery-powered polyimide thin-film resistive heater provides energy, and the device only requires power for a fraction of the total incubation time. The device uses a phase change material (PCM) to regulate temperature to avoid the complexity of a microcontroller. RPA reactions were successfully incubated in 30 minutes using the device.
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用于重组酶聚合酶扩增过程中多个反应温度控制的简单 Poc 装置
对于传染病患者来说,早期准确诊断至关重要。快速诊断可使治疗迅速而有效,并增加完全康复而无并发症的机会。此外,在控制大规模传染病爆发时,如果能够频繁、迅速、经济地对人群进行检测,情况就会大大改善。就特异性和灵敏度而言,核酸扩增检测(NAAT)是诊断传染病的最佳选择之一。聚合酶链反应(PCR)在历史上一直被使用,但复杂的热循环和复杂的 PCR 方案往往限制了 PCR 在临床环境中的应用。由于等温 NAAT 重组酶聚合酶扩增(RPA)更加简便,因此有可能在低资源环境中提供可靠的 POC 诊断。在为等温 NAAT 设计 POC 设备时,创造等温温度条件可能是最大的挑战。这项工作展示了一种灵活而坚固的设备,能够在有利于 POC 检测的条件下培养 3 个 RPA 反应,以同时进行扩增。该装置的建造成本约为 60 美元,易于组装。由电池供电的聚酰亚胺薄膜电阻加热器提供能量,该装置只需一小部分培养时间的电源。该装置使用相变材料(PCM)调节温度,避免了微控制器的复杂性。使用该装置在 30 分钟内成功培养了 RPA 反应。
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