Lateral flow assays (LFA) are quick, simple and cheap assays to analyse a variety of samples at the point of care or in the field, making them one of the most widespread biosensors currently available. They have been successfully employed for the detection of a myriad of different targets (ranging from atoms up to whole cells) in all type of samples (including water, blood, foodstuff and environmental samples). Their operation relies on the capillary flow of the sample within a series of sequential pads with different functionalities aiming to generate a signal indicating the absence/presence (and, in some cases, the concentration) of the analyte of interest. In order to have a user-friendly operation, their development requires the optimization of multiple, interconnected parameters that may overwhelm new developers. In this Tutorial we provide the readers with: 1) the basic knowledge to understand the principles governing an LFA and to take informed decisions during lateral flow strip design and fabrication, 2) a roadmap for optimal LFA development independent of the specific application, 3) a step by step example protocol for the assembly and operation of an LF strip for the detection of Human Immunoglobulin G and 4) an extensive troubleshooting section addressing the most frequent issues in designing, assembling and using LFAs.

Although lateral flow assays (LFA) are currently being used in some point-of-care applications (POC) they cannot still be extended to a broader range of analytes for which higher sensitivities and lower detection limits are required. To overcome such drawbacks, we propose here a simple and facile alternative based on the use of delay hydrophobic barriers fabricated by wax-printing so as to improve the LFA sensitivity. Several wax pillars patterns were printed onto nitrocellulose membrane in order to produce delays as well as pseudo turbulences into the microcapillary flow. The effect of the proposed wax pillar modified devices were also mathematically simulated corroborating the experimental results obtained for the different patterns tested afterwards for detection of HIgG as model protein in a gold nanoparticle-based LFA. The effect of the introduction of such wax-printed pillars was the sensitivity improvement of almost 3-folds in comparison to a conventional free-barrier LFA.

In this article, a novel aptasensor for ochratoxin A (OTA) detection based on a screen-printed carbon electrode (SPCE) modified with polythionine (PTH) and iridium oxide nanoparticles (IrO2 NPs) is presented. The electrotransducer surface is modified with an electropolymerized film of PTH followed by the assembly of IrO2 NPs on which the aminated aptamer selective to OTA is exchanged with the citrate ions surrounding IrO2 NPs via electrostatic interactions with the same surface. Electrochemical impedance spectroscopy (EIS) in the presence of the [Fe(CN)6](-3/-4) redox probe is employed to characterize each step in the aptasensor assay and also for label-free detection of OTA in a range between 0.01 and 100 nM, obtaining one of the lowest limits of detection reported so far for label-free impedimetric detection of OTA (14 pM; 5.65 ng/kg). The reported system also exhibits a high reproducibility, a good performance with a white wine sample, and an excellent specificity against another toxin present in such sample.

The COVID-19 pandemic made clear how our society requires quickly available tools to address emerging healthcare issues. Diagnostic assays and devices are used every day to screen for COVID-19 positive patients, with the aim to decide the appropriate treatment and containment measures. In this context, we would have expected to see the use of the most recent diagnostic technologies worldwide, including the advanced ones such as nano-biosensors capable to provide faster, more sensitive, cheaper, and high-throughput results than the standard polymerase chain reaction and lateral flow assays. Here we discuss why that has not been the case and why all the exciting diagnostic strategies published on a daily basis in peer-reviewed journals are not yet successful in reaching the market and being implemented in the clinical practice.

BackgroundPoint-of-care (POC) tests have become increasingly available and more widely used in recent years. They have been of particular importance to low-income settings, enabling them with clinical capacities that had previously been limited. POC testing programs hold a great potential for significant improvement in low-income health systems. However, as most POC tests are developed in high-income countries, disengagement between developers and end-users inhibit their full potential. This study explores perceptions of POC test end-users in a low-income setting, aiming to support the development of novel POC tests for low-income countries.MethodsA qualitative study was conducted in Mbarara District, Southwestern Uganda, in October 2014. Fifty health care workers were included in seven focus groups, comprising midwives, laboratory technicians, clinical and medical officers, junior and senior nurses, and medical doctors. Discussions were audio-recorded and transcribed verbatim. Transcripts were coded through a data-driven approach for qualitative content analysis.ResultsNineteen different POC tests were identified as currently being in use. While participants displayed being widely accustomed to and appreciative of the use of POC tests, they also assessed the use and characteristics of current tests as imperfect. An ideal POC test was characterized as being adapted to local conditions, thoughtfully implemented in the specific health system, and capable of improving the care of patients. Tests for specific medical conditions were requested. Opinions differed with regard to the ideal distribution of POC tests in the local health system.ConclusionPOC tests are commonly used and greatly appreciated in this study setting. However, there are dissatisfactions with current POC tests and their use. To maximize benefit, stakeholders need to include end-user perspectives in the development and implementation of POC tests. Insights from this study will influence our ongoing efforts to develop POC tests that will be particularly usable in low-income settings.

Iridium oxide nanoparticles (IrO2 NPs) synthesized following a previously reported chemical route are presented as novel tags for immunosensing taking advantage of their electrocatalytic activity towards water oxidation reaction (WOR). Cyclic voltammetry and chronoamperometry for the evaluation of the IrO2 NPs electrocatalytic activity towards WOR at neutral pH were used. The chronoamperometric current recorded at a fixed potential of +1.3 V constituted the analytical signal allowing the quantification of IrO2 NPs at nM levels. Modification of the surface of citrate‐capped IrO2 NPs with anti‐Apolipoprotein E antibodies (αApoE) was successfully achieved and the as‐prepared conjugates were used for the electrocatalytic detection of ApoE Alzheimer disease (AD) biomarker in a magnetosandwich immunoassay, reaching a detection limit of 68 ng/mL. Human plasma of a patient suffering AD was also evaluated, estimating an ApoE concentration of 20 µg/mL which is in concordance with the obtained in previously reported approaches. This novel IrO2 NPs based electrocatalytic assay presents the advantage of the signal generation in the same medium where the immunoassay takes place (PBS, pH 7.4) avoiding the use of additional reagents which also opens the way to future integrated biosensing systems and platforms with interest for other proteins as well as DNA and cells analysis.

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