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低成本、实时聚合酶链反应系统,用于即时医疗诊断。

Low-Cost, Real-Time Polymerase Chain Reaction System for Point-of-Care Medical Diagnosis.

机构信息

Department of Electrical and Computer Engineering, University of Dayton, 300 College Park, Dayton, OH 45469, USA.

Department of Biology, University of Dayton, 300 College Park, Dayton, OH 45469, USA.

出版信息

Sensors (Basel). 2022 Mar 17;22(6):2320. doi: 10.3390/s22062320.

Abstract

Global health crises due to the prevailing Coronavirus Disease 2019 (COVID-19) pandemic have placed significant strain on health care facilities such as hospitals and clinics around the world. Further, foodborne and waterborne diseases are not only spreading faster, but also appear to be emerging more rapidly than ever before and are able to circumvent conventional control measures. The Polymerase Chain Reaction (PCR) system is a well-known diagnostic tool for many applications in medical diagnostics, environmental monitoring, and food and water quality assessment. Here, we describe the design, development, and testing of a portable, low-cost, and real-time PCR system that can be used in emergency health crises and resource-poor situations. The described PCR system incorporates real-time reaction monitoring using fluorescence as an alternative to gel electrophoresis for reaction analysis, further decreasing the need of multiple reagents, reducing sample testing cost, and reducing sample analysis time. The bill of materials cost of the described system is approximately $340. The described PCR system utilizes a novel progressive selective proportional-integral-derivative controller that helps in reducing sample analysis time. In addition, the system employs a novel primer-based approach to quantify the initial target amplicon concentration, making it well-suited for food and water quality assessment. The developed PCR system performed DNA amplification at a level and speed comparable to larger and more expensive commercial table-top systems. The fluorescence detection sensitivity was also tested to be at the same level as commercially available multi-mode optical readers, thus making the PCR system an attractive solution for medical point-of-care and food and water quality assessment.

摘要

由于当前的 2019 年冠状病毒病(COVID-19)大流行,全球卫生危机给世界各地的医院和诊所等医疗机构带来了巨大压力。此外,食源性和水源性疾病不仅传播得更快,而且似乎比以往任何时候都更快地出现,并且能够规避传统的控制措施。聚合酶链反应(PCR)系统是一种众所周知的诊断工具,可用于医学诊断、环境监测以及食品和水质评估等多个应用领域。在这里,我们描述了一种便携式、低成本、实时 PCR 系统的设计、开发和测试,该系统可用于紧急卫生危机和资源匮乏的情况。所描述的 PCR 系统采用实时反应监测,使用荧光作为替代凝胶电泳的反应分析方法,进一步减少了对多种试剂的需求,降低了样品测试成本,并缩短了样品分析时间。该系统的材料成本约为 340 美元。所描述的 PCR 系统利用一种新颖的逐步选择性比例积分微分控制器,有助于缩短样品分析时间。此外,该系统采用了一种新颖的基于引物的方法来定量初始目标扩增子浓度,使其非常适合食品和水质评估。所开发的 PCR 系统在 DNA 扩增水平和速度方面可与更大、更昂贵的商用台式系统相媲美。荧光检测灵敏度也被测试为与市售多模式光学读取器相同,因此使得 PCR 系统成为医疗即时护理和食品及水质评估的有吸引力的解决方案。

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