The development of a multi-disciplinary educational programme in Biomedical Diagnostics - a novel approach.

This paper describes the development of a taught Master’s course in Biomedical Diagnostics using a novel multi-disciplinary approach. This course, the first of its kind in Ireland, covers the science and technology underlying the development of medical diagnostic devices that detect early markers of diseases such as cancer. The ethical impact of these devices on society, the importance of scientific communication, relevant aspects of business entrepreneurial studies and the commercialisation of medical devices are also covered. The course consists of a mixture of theory, intensive laboratory practicals and independent research. The challenges faced in setting up and running the course are described, as well as some of the novel aspects and may provide valuable insight to those involved in the development of high level Masters courses

Detection of benzimidazole carbamates and amino metabolites in liver by surface plasmon resonance-biosensor

This research was funded by the Irish Department of Agriculture, Fisheries and Food under the Food Institutional Research Measure as part of the National Development Plan (Project 05/R&D/TN/355)peer-reviewedTwo surface plasmon resonance (SPR) biosensor screening assays were developed and validated to detect 11 benzimidazole carbamate (BZT) and four amino-benzimidazole veterinary drug residues in liver tissue. The assays used polyclonal antibodies, raised in sheep, to detect BZTs and amino-benzimidazoles. A modified Quick, Easy, Cheap, Effective, Rugged and Safe (QuEChERS) extraction method was developed to isolate benzimidazole carbamate residues. Liver samples were extracted using an acetonitrile extraction method. BZTs were purified by dispersive solid phase extraction (d-SPE) using C18 sorbent. Residues of amino-benzimidazoles were effectively cleaned-up using a simple cyclohexane defatting step. The assays were validated in accordance with the performance criteria described in 2002/657/EC. The BZT assay limit of detection was calculated to be 32 μg kg−1, the detection capability (CCβ) was determined to be 50 μg kg−1 and the mean recovery of analytes was in the range 77–132%. The amino-benzimidazole assay limit of detection was determined to be 41 μg kg−1, the CCβ was determined to be 75 μg kg−1 and analyte recovery was in the range 103–116%. Biosensor assay performance was tested by analysing liver tissue from animals treated with benzimidazole drugs and comparing the results with an ultra high performance liquid chromatography tandem mass spectrometry (UHPLC–MS/MS) confirmatory method. All non-compliant samples were identified using the biosensor assays.Department of Agriculture, Food and the Marin

Hydrophilic polymeric coatings for enhanced, serial-siphon based flow control on centrifugal lab-on-disc platforms

In this paper, we implement rotational flow control on a polymeric microfluidic “lab-on-a-disc” device by combining serial siphoning and capillary valving for sequential release of on-board stored liquid reagents. The functionality of this integrated, multi-step centrifugal assay platform is tightly linked by the capability to establish reproducible, capillary-driven priming of the innately hydrophobic siphon microchannels. We here demonstrate for the first time that spin-coated hydrophilic polymeric films of poly(vinyl alcohol) and (hydroxylpropyl)methylcellulose provide stable contact angles

Comparison of enzyme-linked immunosorbent assay, surface plasmon resonance and biolayer interferometry for screening of deoxynivalenol in wheat and wheat dust

A sample preparation method was developed for the screening of deoxynivalenol (DON) in wheat and wheat dust. Extraction was carried out with water and was successful due to the polar character of DON. For detection, an enzyme-linked immunosorbent assay (ELISA) was compared to the sensor-based techniques of surface plasmon resonance (SPR) and biolayer interferometry (BLI) in terms of sensitivity, affinity and matrix effect. The matrix effects from wheat and wheat dust using SPR were too high to further use this screenings method. The preferred ELISA and BLI methods were validated according to the criteria established in Commission Regulation 519/2014/EC and Commission Decision 2002/657/EC. A small survey was executed on 16 wheat lots and their corresponding dust samples using the validated ELISA method. A linear correlation (r = 0.889) was found for the DON concentration in dust versus the DON concentration in wheat (LOD wheat: 233 g/kg, LOD wheat dust: 458 g/kg)

Development of an autonomous algal toxin analytical platform for aquatic monitoring

Cyclic peptide cyanobacterial toxins, in particular Microcystis aeruginosa, pose a serious health risk to humans and animals alike [1], [2]. Occurring mostly in fresh and brackish water, they have been identified to cause cancer promotion and liver damage [3]. Herein, we describe a portable, microfluidic-based system for in-situ detection of algal toxins in fresh water. The technology development presented here is a fully integrated and portable sample-to-answer centrifugal microfluidics-based system for the detection of toxic cyanobacteria – Microcystin-LR in fresh water. Our unique system employs highly-specific recombinant chicken anti-microcystin antibodies, prepared in-house, with a 3D-printed ‘LASER-photo¬diode’ fluorescent detection technique, also developed in-house. The system has high analytical specificity and sensitivity for detection of toxins below the regulatory limit with intra/inter day coefficient of variation of less than 20%. Dissolvable-film based valving technique was used for flow actuation and integration of multiple assays on the centrifugal cartridge. This new approach forms the basis of a cost efficient, USB-controlled water quality monitoring system. Technically, this integrated system consists of two components; a microfluidic disc (figure 1.A), the disc-holder fabricated and assembled from a 3D-printed casing, with electronic components housed in device. The 5-layered microfluidic disc consists of five reservoirs (figure 1.B), each with a separate venti-lation, aligned radially with inter-connected microchannels. A competitive immunoassay format is utilised to detect free toxin (figure 1.C). Sensitivity, reproducibility and ease-of-use are key features of this monitoring device. The ‘top-down’ optical detection system has been modified for improved detection sensitivity, as well as the elimination of external noise

Point-of-Care Diagnostics in Low Resource Settings: Present Status and Future Role of Microfluidics

The inability to diagnose numerous diseases rapidly is a significant cause of the disparity of deaths resulting from both communicable and non-communicable diseases in the developing world in comparison to the developed world. Existing diagnostic instrumentation usually requires sophisticated infrastructure, stable electrical power, expensive reagents, long assay times, and highly trained personnel which is not often available in limited resource settings. This review will critically survey and analyse the current lateral flow-based point-of-care (POC) technologies, which have made a major impact on diagnostic testing in developing countries over the last 50 years. The future of POC technologies including the applications of microfluidics, which allows miniaturisation and integration of complex functions that facilitate their usage in limited resource settings, is discussed The advantages offered by such systems, including low cost, ruggedness and the capacity to generate accurate and reliable results rapidly, are well suited to the clinical and social settings of the developing world

Electrochemiluminescence platform for the detection of C-reactive proteins : application of recombinant antibody technology to cardiac biomarker detection

This work exploits the high-affinity of recombinant antibodies and low background electrochemiluminescence (ECL) for cardiac-biomarker detection. The developed assay is capable of fg mL-1 detection limits as well as the detection of C-Reactive Protein (CRP) over a clinically relevant range. The assay demonstrated robust reproducibility, selectivity and stability while also highlighting a novel platform for detection of cardiac biomarkers at low concentrations

Fv antibodies to aflatoxin B1 derived from a pre-immunized antibody phage display library system

The production and characterization of recombinant antibodies to aflatoxin B[SUB1] (AFB[SUB1]), a potent mycotoxin and carcinogen is described. The antibody fragments produced were then applied for use in a surface plasmon resonance-based biosensor (BIAcore), which measures biomolecular interactions in 'real-time'. Single chain Fv (scFv) antibodies were generated to aflatoxin B1 from an established phage display system, which incorporated a range of different plasmids for efficient scFv expression. The scFv's were used in the development of a competitive ELISA, and also for the development of surface plasmon resonance (SPR)-based inhibition immunoassays. They were found to be suitable for the detection of AFB[SUB1], in this format, with the assays being sensitive and reproducible