A review of microfabricated electrochemical biosensors for DNA detection

This review article presents an overview of recent work on electrochemical biosensors developed using microfabrication processes, particularly sensors used to achieve sensitive and specific detection of DNA sequences. Such devices are important as they lend themselves to miniaturisation, reproducible mass-manufacture, and integration with other previously existing technologies and production methods. The review describes the current state of these biosensors, novel methods used to produce them or enhance their sensing properties, and pathways to deployment of a complete point-of-care biosensing system in a clinical setting

Development of a rapid, antimicrobial susceptibility test for E. coli based on low-cost, screen-printed electrodes

Antibiotic resistance has been cited by the World Health Organisation (WHO) as one of the greatest threats to public health. Mitigating the spread of antibiotic resistance requires a multipronged approach with possible interventions including faster diagnostic testing and enhanced antibiotic stewardship. This study employs a low-cost diagnostic sensor test to rapidly pinpoint the correct antibiotic for treatment of infection. The sensor comprises a screen-printed gold electrode, modified with an antibiotic-seeded hydrogel to monitor bacterial growth. Electrochemical growth profiles of the common microorganism, Escherichia coli (E. coli) (ATCC 25922) were measured in the presence and absence of the antibiotic streptomycin. Results show a clear distinction between the E. coli growth profiles depending on whether streptomycin is present, in a timeframe of ≈2.5 h (p < 0.05), significantly quicker than the current gold standard of culture-based antimicrobial susceptibility testing. These results demonstrate a clear pathway to a low cost, phenotypic and reproducible antibiotic susceptibility testing technology for the rapid detection of E. coli within clinically relevant concentration ranges for conditions such as urinary tract infections

Biologically modified microelectrode sensors provide enhanced sensitivity for detection of nucleic acid sequences from Mycobacterium tuberculosis

This paper describes improved sensitivity when using biosensors based on microfabricated microelectrodes to detect DNA, with the goal of progressing towards a low cost and mass manufacturable assay for antibiotic resistance in tuberculosis (TB). The microelectrodes gave a near 20 times improvement in sensitivity compared to polycrystalline macroelectrodes. In addition, experimental parameters such as redox mediator concentration and experimental technique were investigated and optimised. It was found that lower concentrations of redox mediator gave higher signal changes when measuring hybridisation events and, at these lower concentrations, square wave voltammetry was more sensitive and consistent than differential pulse voltammetry. Together, this paper presents a quantifiable comparison of macroelectrode and microelectrode DNA biosensors. The final assay demonstrates enhanced sensitivity through reduction of sensor size, reduction of redox mediator concentration and judicious choice of detection technique, therefore maintaining manufacturability for incorporation into point of care tests and lab-on-a-chip devices

Development and Optimization of Durable Microelectrodes for Quantitative Electroanalysis in Molten Salt

Microfabricated square electrodes with finely controlled highly reproducible dimensions have been developed for electrochemical analysis of high-temperature molten salt (MS). These microelectrodes have been fabricated using photolithographic techniques on silicon wafers and have been designed for operation in lithium chloride/potassium chloride eutectic salt at and ~500 °C. The electrodes are constructed from a series of patterned layers, and their development has involved a systematic study and optimization of a number of different material combinations. This has resulted in a process for making electrodes that represents a step change in capability, delivering the first robust microelectrode device capable of quantitative electroanalysis in a MS system at 500 °C

Test structure and measurement system for characterising the electrochemical performance of nanoelectrode structures

This paper presents a complete test structure and characterisation system for the evaluation of nanoelectrode technology. It integrates microfabricated nanoelectrodes for electrochemical measurements, 3D printing and surface tensionconfined microfluidics. This system exploits the inherent analytical advantages of nanoelectrodes that enables their operation with small volume samples, which has potential applications for onwafer measurements

Optimization of Nafion Polymer Electrolyte Membrane Design and Microfabrication

Nafion is a solid electrolyte polymer that can be used as a sensor membrane in microfabricated electrochemical oxygen sensors. It allows ions to be transported between the sensor electrodes and removes the need for a liquid electrolyte. Here we used a series of small square Nafion test structures, fabricated on a variety of materials using standard thin-film patterning techniques, to optimize the design and processing of Nafion membranes. Measurements showed that the choice of photoresist developer is critical. Use of diluted MF-26A developer provided the most effective and manufacturable process. The underlying material also had an influence on robustness, with silicon dioxide and platinum giving the longest membrane lifetime under simulated conditions of use. Membrane size had no clear effect on lifetime, and under optimal processing conditions there were minimal failures even under continuous mechanical agitation for up to six weeks. We also developed test electrodes covered by Nafion, and showed that they were effective at supporting electrochemical oxygen detection

In vivo validation of a miniaturized electrochemical oxygen sensor for measuring intestinal oxygen tension

Recent advances in the fields of electronics and microfabrication techniques have led to the development of implantable medical devices for use within the field of precision medicine. Monitoring visceral surface tissue O2 tension (ptO2) by means of an implantable sensor is potentially useful in many clinical situations including the peri-operative management of patients undergoing intestinal resection and anastomosis. This concept could provide a means by which treatment could be tailored to individual patients. This study describes the in vivo validation of a novel miniaturised electrochemical O2 sensor to provide real-time data on intestinal ptO2. A single O2 sensor was placed onto the serosal surface of the small intestine of anaesthetised rats that were exposed to ischaemic (superior mesenteric artery occlusion) and hypoxaemic (alterations in inspired fractional O2 concentrations) insults. Control experiments demonstrated that the sensors function and remain stable in an in vivo environment. Intestinal ptO2 decreased following superior mesenteric artery occlusion and with reductions in inspired O2 concentrations. These results were reversible after reinstating blood flow or increasing inspired O2 concentrations. We have successfully developed an anaesthetised rat intestinal ischaemic and hypoxic model for validation of a miniaturised O2 sensor to provide real-time measurement of intestinal ptO2. Our results support further validation of the sensors in physiological conditions using a large animal model to provide evidence of their use in clinical applications where monitoring visceral surface tissue O2 tension is important

An electrochemical SARS-CoV-2 biosensor inspired by glucose test strip manufacturing processes

Accurate and rapid diagnostic tests are critical to reducing the impact of SARS-CoV-2. This study presents early, but promising measurements of SARS-CoV-2 using the ACE2 enzyme as the recognition element to achieve clinically relevant detection. The test provides a scalable route to sensitive, specific, rapid and low cost mass testing