No abstract

Isothermal nucleic acid amplification tests (NAAT) in a Lab-on-a-Chip (LoC) format promise to bring high-accuracy, non-instrumented rapid tests to the point of care. Reliable rapid tests for infectious diseases allow for early diagnosis and treatment, which in turn enables better containment of potential outbreaks and fewer complications. A critical component to LoC NAATs is the heating element, as all NAAT protocols require incubation at elevated temperatures. We propose a cheap, integrated, self-regulating resistive heating solution that uses 2xAAA alkaline batteries as the power source, can maintain temperatures in the 60–63°C range for at least 25 minutes, and reaches the target range from room temperature in 5 minutes. 4 heating element samples with different electrical characteristics were evaluated in a thermal mock-up for a LoC NAAT device. An optimal heating element candidate was chosen based on temperature profiling. The optimal candidate was further evaluated by thermal modelling via finite element analysis of heat transfer and demonstrated suitable for isothermal nucleic acid amplification.

In recent years, additive manufacturing has steadily gained attention in both research and industry. Applications range from prototyping to small-scale production, with 3D printing offering reduced logistics overheads, better design flexibility and ease of use compared with traditional fabrication methods. In addition, printer and material costs have also decreased rapidly. These advantages make 3D printing attractive for application in microfluidic chip fabrication. However, 3D printing microfluidics is still a new area. Is the technology mature enough to print complex microchannel geometries, such as droplet microfluidics? Can 3D-printed droplet microfluidic chips be used in biological or chemical applications? Is 3D printing mature enough to be used in every research lab? These are the questions we will seek answers to in our systematic review. We will analyze (1) the key performance metrics of 3D-printed droplet microfluidics and (2) existing biological or chemical application areas. In addition, we evaluate (3) the potential of large-scale application of 3D printing microfluidics. Finally, (4) we discuss how 3D printing and digital design automation could trivialize microfluidic chip fabrication in the long term. Based on our analysis, we can conclude that today, 3D printers could already be used in every research lab. Printing droplet microfluidics is also a possibility, albeit with some challenges discussed in this review.

Non-instrumented nucleic acid amplification tests (NINAAT) are a novel paradigm in portable molecular diagnostics. They offer the high detection accuracy characteristic of nucleic acid amplification tests (NAAT) in a self-contained device, without the need for any external instrumentation. These Point-of-Care tests typically employ a Lab-on-a-Chip for liquid handling functionality, and perform isothermal nucleic acid amplification protocols that require low power but high accuracy temperature control in a single well-defined temperature range. We propose temperature control solutions based on commercially available heating elements capable of meeting these challenges, as well as demonstrate the process by which such elements can be fitted to a NINAAT system. Self-regulated and thermostat-controlled resistive heating elements were evaluated through experimental characterization as well as thermal analysis using the finite element method (FEM). We demonstrate that the proposed solutions can support various NAAT protocols, as well as demonstrate an optimal solution for the loop-mediated isothermal amplification (LAMP) protocol. Furthermore, we present an Arduino-compatible open-source thermostat developed for NINAAT applications.

We present a finite element model for the simulation of the resistive heating of microchannels in disposable molecular diagnostics devices by means of various resistive heating elements, and demonstrate the validity of this model through experiments. Polyimide etched foil heaters and heaters based on positive temperature coefficient (PTC) ceramics could be simple, cost-efficient and robust means of heating disposable Lab-on-a-Chip devices that depend on maintaining a given temperature range in the channels for an extended time (i.e. 15-60 minutes) with a precision of ±1-5°C. However, the design of these devices is a slow and costly process due to the many design factors involved. We demonstrate a finite element model that could reduce design time and save on prototyping costs. The validity of the model is demonstrated through various PMMA test structures designed to emulate a Lab-on-a-Chip device capable of supporting isothermal nucleic acid amplification reactions. With our experimental setups we were able to produce and maintain target temperatures for over 45 minutes with a precision of at most ±1°C deviation from the set point in a battery-operated test system without the use of a thermostat. The physical parameters of the available resistive heating elements were used in our finite element model, and the results of the simulations compared to experimental data.