Low-Cost, Real-Time Polymerase Chain Reaction System with Integrated RNA Extraction.

Rapid, easy-to-use, and low-cost systems for biological sample testing are important for point-of-care diagnostics and various other health applications. The recent pandemic of Coronavirus Disease 2019 (COVID-19) caused by the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) showed an urgent need to rapidly and accurately identify the genetic material of SARS-CoV-2, an enveloped ribonucleic acid (RNA) virus, in upper respiratory specimens from people. In general, sensitive testing methods require genetic material extraction from the specimen. Unfortunately, current commercially available extraction kits are expensive and involve time-consuming and laborious extraction procedures. To overcome the difficulties associated with common extraction methods, we propose a simple enzymatic assay for the nucleic acid extraction step using heat mediation to improve the polymerase chain reaction (PCR) reaction sensitivity. Our protocol was tested on Human Coronavirus 229E (HCoV-229E) as an example, which comes from the large coronaviridae family of viruses that affect birds, amphibians, and mammals, of which SARS-CoV-2 is a member. The proposed assay was performed using a low-cost, custom-made, real-time PCR system that incorporates thermal cycling and fluorescence detection. It had fully customizable reaction settings to allow versatile biological sample testing for various applications, including point-of-care medical diagnosis, food and water quality testing, and emergency health situations. Our results show that heat-mediated RNA extraction is a viable extraction method when compared to commercial extraction kits. Further, our study showed that extraction has a direct impact on purified laboratory samples of HCoV-229E, but no direct impact on infected human cells. This is clinically relevant, as it allows us to circumvent the extraction step on clinical samples when using PCR.

Experimental Evaluation of Sensor Fusion of Low-Cost UWB and IMU for Localization under Indoor Dynamic Testing Conditions.

Autonomous systems usually require accurate localization methods for them to navigate safely in indoor environments. Most localization methods are expensive and difficult to set up. In this work, we built a low-cost and portable indoor location tracking system by using Raspberry Pi 4 computer, ultra-wideband (UWB) sensors, and inertial measurement unit(s) (IMU). We also developed the data logging software and the Kalman filter (KF) sensor fusion algorithm to process the data from a low-power UWB transceiver (Decawave, model DWM1001) module and IMU device (Bosch, model BNO055). Autonomous systems move with different velocities and accelerations, which requires its localization performance to be evaluated under diverse motion conditions. We built a dynamic testing platform to generate not only the ground truth trajectory but also the ground truth acceleration and velocity. In this way, our tracking system's localization performance can be evaluated under dynamic testing conditions. The novel contributions in this work are a low-cost, low-power, tracking system hardware-software design, and an experimental setup to observe the tracking system's localization performance under different dynamic testing conditions. The testing platform has a 1 m translation length and 80 µm of bidirectional repeatability. The tracking system's localization performance was evaluated under dynamic conditions with eight different combinations of acceleration and velocity. The ground truth accelerations varied from 0.6 to 1.6 m/s2 and the ground truth velocities varied from 0.6 to 0.8 m/s. Our experimental results show that the location error can reach up to 50 cm under dynamic testing conditions when only relying on the UWB sensor, with the KF sensor fusion of UWB and IMU, the location error decreases to 13.7 cm.

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

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.