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Microbial fuel cells (MFCs) are an ecologically friendly technology that can recover electricity and simultaneously treat wastewater. Among all the influential factors, cathode material and catalyst play a crucial role in electricity production and oxygen reduction. In this study, Pt nanocatalysts deposited on single-walled carbon nanotubes (Pt/SWCNTs) were synthesized by the deposition-precipitation (DP) method under optimal conditions. The results show that DP might be a promising method for the preparation of Pt/SWCNTs due to its simple, cost-effective and time-saving procedure, in addition to being highly efficient at creating small Pt particles (0.9 nm) that were very uniformly distributed. The synthesized Pt/SWCNTs suspension was spray-coated on to carbon cloth and then used as a cathode for MFCs. The electricity generation of MFCs equipped with a Pt/SWCNTs cathode was evaluated in terms of open circuit voltage (0.6954 V), internal resistance (63.3 Ω⋅m²) and maximum power density (2,022 mW/m²). The chemical oxygen demand removal, biological oxygen demand removal, total dissolved solids removal, total suspended solids removal and silver recovery was satisfactory at 84.5%, 74.0%, 45.7%, 60.0% and 99.0%, respectively. Therefore, Pt/SWCNTs from the DP method was identified as a potential candidate to replace commercial Pt-carbon cloth for MFC cathodes. [ABSTRACT FROM AUTHOR]

Two microfluidic systems have been developed for specific analysis of L-glutamate in food based on substrate recycling fluorescence detection. L-glutamate dehydrogenase and a novel enzyme, D-phenylglycine aminotransferase, were covalently immobilized on (i) the surface of silicon microchips containing 32 porous flow channels of 235 μm depth and 25 μm width and (ii) polystyrene Poros™ beads with a particle size of 20 μm. The immobilized enzymes recycle L-glutamate by oxidation to 2-oxoglutarate followed by the transfer of an amino group from D-4-hydroxyphenylglycine to 2-oxoglutarate. The reaction was accompanied by reduction of nicotinamide adenine dinucleotide (NAD+) to NADH, which was monitored by fluorescence detection ([variant_greek_epsilon]ex=340 nm, [variant_greek_epsilon]em=460 nm). First, the microchip-based system, L-glutamate was detected within a range of 3.1–50.0 mM. Second, to be automatically determined, sequential injection analysis (SIA) with the bead-based system was investigated. The bead-based system was evaluated by both flow injection analysis and SIA modes, where good reproducibility for L-glutamate calibrations was obtained (relative standard deviation of 3.3% and 6.6%, respectively). In the case of SIA, the beads were introduced and removed from the microchip automatically. The immobilized beads could be stored in a 20% glycerol and 0.5 mM ethylenediaminetetraacetic acid solution maintained at a pH of 7.0 using a phosphate buffer for at least 15 days with 72% of the activity remaining. The bead-based system demonstrated high selectivity, where L-glutamate recoveries were between 91% and 108% in the presence of six other L-amino acids tested. [ABSTRACT FROM AUTHOR]

A paper-based potentiometric sensor integrated with a polymeric hydrogel has been developed for sodium ion (Na + ) determination in human urine. The construction of an all-solid-state ion selective electrode (s-ISE) and an all-solid-state reference electrode (s-RE) on a photo paper substrate was achieved using an inkjet printing method. For s-ISE fabrication, carbon nanotubes (CNTs) and gold nanoparticles (AuNPs) were printed on the substrate as a nanocomposite solid contact. A polymeric hydrogel containing lithium acetate (CH 3 COOLi) was then prepared and used as an intermediate layer to improve the adhesion between the ion selective membrane (ISM) and the AuNP/CNT solid contact, leading to enhanced detection sensitivity. The printed s-RE consisted of a pseudo silver/silver chloride electrode (p-Ag/AgCl) coated with a polymeric hydrogel containing KCl to improve the potential stability of the sensor. Under the optimal conditions, the hydrogel-integrated paper-based potentiometric sensor provided a response toward Na + over a linear range of 10 -7 M to 1 M with a near Nernstian slope of 56.42 ± 0.68 mV per decade. This sensor exhibited fast response, good sensitivity, and reasonable selectivity for Na + measurement. Furthermore, the developed sensor was effectively applied for the detection of Na + in urine samples with high accuracy. The presented work can be considered as a good addition to the growing field of potentiometric analytical platforms suitable for large-scale production using inkjet printing technology.

Monitoring various biomarkers in saliva samples emerges as a dynamic and non-invasive method. However, the high viscosity of saliva presents a distinct challenge when integrating paper-based platforms for on-site analysis. In addressing this challenge, we introduced the capillary-driven microfluidic paper-based analytical devices (μCD-PAD) designed for user-friendly and simultaneous detection of ethanol and tetrahydrocannabinol (THC) in saliva without a sample preparation step. Employing a colorimetric approach, we quantified both analytes. Synthetic salivas of varying viscosity flowed seamlessly to the detection zone without needing a sample preparation step, and no impact on colorimetric detection due to viscosity was observed (RSD <5 %). Within 10 min after the solution reached the detection zone, the device produced a homogeneous color signal, easily analyzed by a smartphone camera. To extend the application for determination to cover a legal limit concentration of ethanol and concentration of salivary THC even 24 h after marijuana consumption, the detection time of 30 min was optimized. Moreover, a saliva sample containing both analytes was used to demonstrate the capability of the developed device to detect ethanol and THC simultaneously. No cross-talk between ethanol and THC occurred and showed recovery in the 98-102 % for ethanol and 95-105 % for THC with acceptable accuracy. This developed device exhibits excellent potential for forensic applications, providing a user-friendly, cost-effective, and real-time screening tool for detecting ethanol and THC in saliva., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

The similar transmission patterns and early symptoms of respiratory viral infections, particularly severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), influenza (H1N1), and respiratory syncytial virus (RSV), pose substantial challenges in the diagnosis, therapeutic management, and handling of these infectious diseases. Multiplexed point-of-care testing for detection is urgently needed for prompt and efficient disease management. Here, we introduce an electrochemical paper-based analytical device (ePAD) platform for multiplexed and label-free detection of SARS-CoV-2, H1N1, and RSV infection using immobilized pyrrolidinyl peptide nucleic acid probes. Hybridization between the probes and viral nucleic acid targets causes changes in the electrochemical response. The resulting sensor offers high sensitivity and low detection limits of 0.12, 0.35, and 0.36 pM for SARS-CoV-2 (N gene), H1N1, and RSV, respectively, without showing any cross-reactivities. The amplification-free detection of extracted RNA from 42 nasopharyngeal swab samples was successfully demonstrated and validated against reverse-transcription polymerase chain reaction (range of cycle threshold values: 17.43-25.89). The proposed platform showed excellent clinical sensitivity (100 %) and specificity (≥97 %) to achieve excellent agreement (κ ≥ 0.914) with the standard assay, thereby demonstrating its applicability for the screening and diagnosis of these respiratory diseases., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

A novel approach for simultaneous detection of iron and potassium via a smartphone-based potentiometric method is proposed in this study. The screen printed electrodes were modified with carbon black nanomaterial and ion selective membrane including zinc (II) phtalocyanine as the ionophore. The developed Fe 3+ -selective electrode and K + -selective electrode exhibited detection limits of 1.0 × 10 -6  M and 1.0 × 10 -5  M for Fe 3+ and K + ions, respectively. The electrodes were used to simultaneously detect Fe 3+ and K + ions in apple juice, skim milk, soybean and coconut water samples with recovery values between 90%-100.5%, and validated against inductively coupled plasma-optical emission spectrometry. Due to the advantageous characteristics of the sensors and the portability of Near Field Communication potentiometer supported with a smartphone application, the proposed method offers sensitive and selective detection of iron and potassium ions in food and beverage samples at the point of need., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests. Tugba Ozer reports financial support was provided by TUBITAK under project number 122Z721. Tugba Ozer reports a relationship with TUBITAK that includes: funding grants. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier Ltd.)

A novel colorimetric method utilizing a bi-enzyme assay using silver nanoplates (AgNPls) as a direct signal source was developed to enable rapid insecticide detection. This innovative system leverages the in situ generated H 2 O 2 from the consecutive enzyme-catalyzed reactions of acetylcholine hydrolysis and choline oxidation to introduce oxidative etching of AgNPls, transforming them into aggregated silver nanospheres (AgNSs). The morphological transformation of silver nanoparticles could be observed with the naked eye due to the solution's color shifts from pink-violet to blue-violet. The presence of insecticide, i.e. , dichlorvos (DDVP), could inhibit acetylcholinesterase activity, thereby limiting H 2 O 2 production and affecting the transformation of AgNPls into aggregated AgNSs. Furthermore, the extent of AgNPl-to-aggregated AgNS transformation and the subsequent solution's color change was inversely proportional to the amount of DDVP. Under optimal conditions, the developed bi-enzyme assay enables the quantification of DDVP within 5 minutes, achieving detection limits of 0.5 ppm and 0.1 ppm by naked-eye detection and UV-visible spectrophotometry, respectively. Furthermore, the practical application of this assay was validated for detecting insecticides in real vegetable samples, demonstrating both accuracy and reliability., Competing Interests: There are no conflicts to declare., (This journal is © The Royal Society of Chemistry.)

A portable device offering effortlessness, mobility, and affordability for real-time and on-site monitoring of heavy metals is currently in great demand to maintain environmental sustainability. Herein, a platform utilizing a biopolymeric gel-based electrolyte for the on-field simultaneous determination of Cd(II) and Pb(II) is described. Pectin, a natural polymer, was exploited as a chemical delivery medium on account of its biodegradability, environmental friendliness, and rapid dissolving characteristics. The gel electrolyte was prepared by having pectin dissolved in KCl mixed with Sb(III)-Bi(III) bimetallic alloy solution, and casted onto a paper substrate. An in situ bimetallic alloy and pre-mixed bismuth nanoparticles modified screen-printed graphene electrode (Sb-Bi/BiNP/SPGE) were employed to enhance the electrochemical signals of Cd(II) and Pb(II) for the differential pulse anodic stripping voltammetry (DPASV). It was demonstrated that the platform was capable of generating sharp and well-defined current signals, achieving the low detection limits of 50.98 ng mL -1 for Cd(II) and 40.80 ng mL -1 for Pb(II). The reproducibility, as indicated by the relative standard deviation, was found to be less than 10.4 % (n = 10) for the developed gel-based device when coupled with a wireless near field communication (NFC) potentiostat. Lastly, the obtained sensor was applied for quantification of Cd and Pb in potentially contaminated groundwater samples. The recoveries obtained were satisfactorily within the acceptable range. The newly designed platform exhibited several advantages, including small sample volume (μL), low-cost, no sample preparation requirements, and being environmentally friendly. The convenience of a portable device utilizing the proposed biopolymeric gel-based electrolyte for on-field analysis makes it highly appealing for various applications., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Laser-induced graphene (LIG) electrodes have become popular for electrochemical sensor fabrication due to their simplicity for batch production without the use of reagents. The high surface area and favorable electrocatalytic properties also enable the design of small electrochemical devices while retaining the desired electrochemical performance. In this work, we systematically investigated the effect of LIG working electrode size, from 0.8 mm to 4.0 mm diameter, on their electrochemical properties, since it has been widely assumed that the electrochemistry of LIG electrodes is independent of size above the microelectrode size regime. The background and faradaic current from cyclic voltammetry (CV) of an outer-sphere redox probe [Ru(NH3)6] 3+ showed that smaller LIG electrodes had a higher electrode roughness factor and electroactive surface ratio than those of the larger electrodes. Moreover, CV of the surface-sensitive redox probes [Fe(CN)6] 3- and dopamine revealed that smaller electrodes exhibited better electrocatalytic properties, with enhanced electron transfer kinetics. Scanning electron microscopy, Raman spectroscopy, and X-ray photoelectron spectroscopy showed that the physical and chemical surface structure were different at the electrode center versus the edges, so the electrochemical properties of the smaller electrodes were improved by having rougher surface and more density of the graphitic edge planes, and more oxide-containing groups, leading to better electrochemistry. The difference could be explained by the different photothermal reaction time from the laser scribing process that causes different stable carbon morphology to form on the polymer surface. Our results give a new insight on relationships between surface structure and electrochemistry of LIG electrodes and are useful for designing miniaturized electrochemical devices., Competing Interests: Declaration of interests The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Following the global COVID-19 pandemic triggered by SARS-CoV-2, the need for rapid, specific and cost-effective point-of-care diagnostic solutions remains paramount. Even though COVID-19 is no longer a public health emergency, the disease still poses a global threat leading to deaths, and it continues to change with the risk of new variants emerging causing a new surge in cases and deaths. Here, we address the urgent need for rapid, cost-effective and point-of-care diagnostic solutions for SARS-CoV-2. We propose a multiplexed DNA-based sensing platform that utilizes inkjet-printed nanostructured gold electrodes and an inkjet-printed battery-free near-field communication (NFC) potentiostat for the simultaneous quantitative detection of two SARS-CoV-2 genes, the ORF1ab and the N gene. The detection strategy based on the formation of an RNA-DNA sandwich structure leads to a highly specific electrochemical output. The inkjet-printed nanostructured gold electrodes providing a large surface area enable efficient binding and increase the sensitivity. The inkjet-printed battery-free NFC potentiostat enables rapid measurements and real-time data analysis via a smartphone application, making the platform accessible and portable. With the advantages of speed (5 min), simplicity, sensitivity (low pM range, ∼450% signal gain) and cost-effectiveness, the proposed platform is a promising alternative for point-of-care diagnostics and high-throughput analysis that complements the COVID-19 diagnostic toolkit., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Tuberculosis (TB) currently remains a major life-threatening disease as it can be fatal if not treated properly or in a timely manner. Herein, we first describe a disposable and cost-effective paper-based electrochemical biosensor based on a gold particle-decorated carboxyl graphene (AuPs/GCOOH)-modified electrode for detecting heat shock protein (Hsp16.3), which is a specific biomarker indicating the onset of TB infection. The device pattern was first engineered to facilitate detection procedures and printed on low-cost filter paper to create hydrophobic and hydrophilic regions using a wax printing technique. Immunoassays proceeded in a half-sandwich format because it is a reagent-less approach and requires no labeling step. The fabrication of the immunosensor began with GCOOH drop casting, the electrochemical deposition of AuPs, and the establishment of a biorecognition layer against Hsp16.3 utilizing 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC)/N-hydroxysuccinimide (NHS)-sulfo standard chemistry. The appearance of Hsp16.3 resulted in a substantial decrease in the electrochemical signal response of the redox probe employed [Fe (CN) 6 ] 3-/4- due to the created immunocomplexes that possess insulation properties. GCOOH enables direct antibody immobilization, and AuPs enhance the electrochemical properties of the sensor. This proposed immunosensor, while requiring only a miniscule sample volume (5 μL), achieved superior performance in terms of the limit of detection, measuring at 0.01 ng/mL. Our platform was confirmed to be highly specific to Hsp16.3 and can rapidly detect TB-infected sera without necessitating any pre-enrichment (20 min), making it an alternative and particularly suitable for the early diagnosis of TB in resource-scarce countries., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

This review explores the advancements in nanomaterial-based electrochemical sensors for the multiplex detection of medicinal compounds. The growing demand for efficient and selective detection methods in the pharmaceutical field has prompted significant research into the development of electrochemical sensors employing nanomaterials. These materials, defined as functional materials with at least one dimension between 1 and 100 nanometers, encompass metal nanoparticles, polymers, carbon-based nanocomposites, and nano-bioprobes. These sensors are characterized by their enhanced sensitivity and selectivity, playing a crucial role in simultaneous detection and offering a comprehensive analysis of multiple medicinal complexes within a single sample. The review comprehensively examines the design, fabrication, and application of nanomaterial- based electrochemical sensors, focusing on their ability to achieve multiplex detection of various medicinal substances. Insights into the strategies and nanomaterials employed for enhancing sensor performance are discussed. Additionally, the review explores the challenges and future perspectives of this evolving field, highlighting the potential impact of nanomaterial-based electrochemical sensors on the advancement of medicinal detection technologies., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

An automated microfluidic electrochemical platform was developed for the rapid in-field analysis of arsenic speciation. Herein, we integrated an electrochemical sensing and microfluidic channel for the simultaneous determination of As(III) and total inorganic As (total iAs) within a single device. The platform was fabricated by assembling a gold nanoparticle-modified screen-printed graphene electrode (AuNP/SPGE) on a hydrophilic polyethylene terephthalate (PET) sheet that was specially designed to enclose a microfluidic channel with dual flow channels for separate determination of the two species. While As(III) can be promptly detected with the AuNP/SPGE on one end, thioglycolic acid stored in glass fiber is employed on the other end to reduce As(V) before being electrochemically analyzed on the AuNP/SPGE as total iAs; the difference represents the amount of As(V). With a wireless potentiostat and a smartphone equipped with Bluetooth technology, the overall procedure can be fully automated and accomplished merely within 9 min. The linear ranges for the determination of As(III) and total iAs were found to be 50-1000 and 100-1500 ng/mL with detection limits of 3.7 and 17 ng/mL, respectively. The proposed method was validated and applied for the inorganic As speciation of various food samples with satisfactory results compared to those obtained with the standard HPLC-ICP‒MS protocol. This novel microfluidic electrochemical platform offers numerous advantages, notably for its simplicity, speed, low cost, and portability for on-site analysis, which conclusively makes it a highly promising analytical device for the speciation of inorganic arsenic., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper, (Copyright © 2023 Elsevier B.V. All rights reserved.)

The coronavirus disease 2019 (COVID-19) pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been a significant health issue globally. Point-of-care (POC) testing that can offer a rapid and accurate diagnosis of SARS-CoV-2 at the early stage of infection is highly desirable to constrain this outbreak, especially in resource-limited settings. Herein, we present a G-quadruplex DNAzyme-based electrochemical assay that is integrated with a sequential flow controllable microfluidic device for the detection of SARS-CoV-2 cDNA. According to the detection principle, a pyrrolidinyl peptide nucleic acid probe is immobilized on a screen-printed graphene electrode for capturing SARS-CoV-2 DNA. The captured DNA subsequently hybridizes with another DNA probe that carries a G-quadruplex DNAzyme as the signaling unit. The G-quadruplex DNAzyme catalyzes the H 2 O 2 -mediated oxidation of hydroquinone to benzoquinone that can be detected using square-wave voltammetry to give a signal that corresponds to the target DNA concentration. The assay exhibited high selectivity for SARS-CoV-2 DNA and showed a good experimental detection limit at 30 pM. To enable automation, the DNAzyme-based assay was combined with a capillary-driven microfluidic device featuring a burst valve technology to allow sequential sample and reagent delivery as well as the DNA target hybridization and enzymatic reaction to be operated in a precisely controlled fashion. The developed microfluidic device was successfully applied for the detection of SARS-CoV-2 from nasopharyngeal swab samples. The results were in good agreement with the standard RT-PCR method and could be performed within 20 min. Thus, this platform offers desirable characteristics that make it an alternative POC tool for COVID-19 diagnosis.

Sepsis, an infectious disease affecting millions of people's health worldwide each year, calls for urgent attention to an improvement of analytical devices. Chemiluminescence immunoassay is a typical diagnostic method utilized to assess the risk development of sepsis. However, due to its high-cost, delayed, and complicated procedure, the practical utilization is therefore undoubtedly limited, especially for point-of-care test. Herein, we fabricated for the first time an immunosensor based on dendritic copper nanostructures (CuNSs) combined with 4-aminobenzoic acid (4-AB, the diazonium salt) as antibody linker modified on a screen-printed graphene electrode for the early detection of the sepsis biomarker interleukin-6 (IL-6). The electrode fabrication is made by electrodeposition, thus eliminating the multistep of nanomaterial synthesis and time wasting. The resulting dendritic CuNSs significantly increase the effective surface area (1.2 times) and the sensor's performance. The morphology of this combination was characterized using CV, EIS, SEM, EDX, and FTIR techniques. In the detection process, the appearance of IL-6 suppresses the current response of the redox probe indicator measured by differential pulse voltammetry due to the antibody-antigen complex. The subtraction of signal (ΔI) was interpreted as IL-6 concentration. This sensor exhibited a linear range from 0.05 to 500 pg mL -1 with low detection limit of 0.02 pg mL -1 , proving a possibility for early sepsis screening. In addition, the established immunosensor can successfully quantify IL-6 in human serum sample, in which the results agreed well with those achieved using the standard approach, further showing high practical applicability of this developed immunosensor., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

Electrochemical DNA sensors can be operated in either static or flow-based detection schemes. In static schemes, manual washing steps are still necessary, resulting in a tedious and time-consuming process. In contrast, in flow-based electrochemical sensors, the current response is collected when the solution flows through the electrode continuously. However, the drawback of such a flow system is the low sensitivity due to the limited time for the interaction between the capturing element and the target. Herein, we propose a novel electrochemical capillary-driven microfluidic DNA sensor to combine the advantages of static and flow-based electrochemical detection systems into a single device by incorporating burst valve technology. The microfluidic device with a two-electrode configuration was applied for the simultaneous detection of two different DNA markers, human immunodeficiency virus-1 (HIV-1) and hepatitis C virus (HCV) cDNA, via the specific interaction between pyrrolidinyl peptide nucleic acids (PNA) probes and the DNA target. The integrated system, while requiring a small sample volume (7 μL for each sample loading port) and less analysis time, achieved good performance in terms of the limits of detection (LOD) (3SD blank /slope) and quantification (LOQ) (10SD blank /slope) at 1.45 nM and 4.79 nM for HIV and 1.20 nM and 3.96 nM for HCV, respectively. The simultaneous detection of HIV-1 and HCV cDNA prepared from human blood samples showed results that are in complete agreement with the RT‒PCR assay. The results qualify this platform as a promising alternative for the analysis of either HIV-1/HCV or coinfection that can be easily adapted for other clinically important nucleic acid-based markers., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Amino acid conductive polymers can easily form a thin film on a sensor surface by an electrochemical process. Therefore, we are pioneers in reporting the electropolymerization of L-methionine on the surface of a screen-printed graphene electrode to obtain a disposable electrochemical sensor for determining drug metabolites (5-aminosalicylic acid (5-ASA) and sulfapyridine (SPD)) of sulfasalazine (SSZ) simultaneously. In this work, the developed sensor was facilely created through a single step of electropolymerization under mild conditions (0.1 M phosphate buffer pH 7.0) using cyclic voltammetry. Important parameters in the synthesis process were systematically investigated followed by surface composition and morphology studies. Then, analytical performances, comprising sensitivity, selectivity, stability, reproducibility, and sample preparation, were carefully evaluated. Under optimal conditions, the proposed methodology demonstrated a highly sensitive and selective simultaneous detection of 5-ASA and SPD with wide linear dynamic ranges of 1-50 μM and 80-250 μM and low detection limits of 0.60 and 0.57 μM for 5-ASA and SPD, respectively. To evaluate the potential of the designed sensor, it was successfully applied by simultaneously determining 5-ASA and SPD in real human urine samples on the same day (intra-day study) and on three different days (inter-day study).

A capillary-driven microfluidic sequential flow device, designed for eventual at-home or doctor's office use, was developed to perform an enzyme-linked immunosorbent assay (ELISA) for serology assays. Serology assays that detect SARS-CoV-2 antibodies can be used to determine prior infection, immunity status, and/or individual vaccination status and are typically run using well-plate ELISAs in centralized laboratories, but in this format SARs-CoV-2 serology tests are too expensive and/or slow for most situations. Instead, a point-of-need device that can be used at home or in doctor's offices for COVID-19 serology testing would provide critical information for managing infections and determining immune status. Lateral flow assays are common and easy to use, but lack the sensitivity needed to reliably detect SARS-CoV-2 antibodies in clinical samples. This work describes a microfluidic sequential flow device that is as simple to use as a lateral flow assay, but as sensitive as a well-plate ELISA through sequential delivery of reagents to the detection area using only capillary flow. The device utilizes a network of microfluidic channels made of transparency film and double-sided adhesive combined with paper pumps to drive flow. The geometry of the channels and storage pads enables automated sequential washing and reagent addition steps with two simple end-user steps. An enzyme label and colorimetric substrate produce an amplified, visible signal for increased sensitivity, while the integrated washing steps decrease false positives and increase reproducibility. Naked-eye detection can be used for qualitative results or a smartphone camera for quantitative analysis. The device detected antibodies at 2.8 ng mL -1 from whole blood, while a well-plate ELISA using the same capture and detection antibodies could detect 1.2 ng mL -1 . The performance of the capillary-driven immunoassay (CaDI) system developed here was confirmed by demonstrating SARS-CoV-2 antibody detection, and we believe that the device represents a fundamental step forward in equipment-free point-of-care technology.

A significant bottleneck exists for mass-production of ion-selective electrodes despite recent developments in manufacturing technologies. Here, we present a fully-automated system for large-scale production of ISEs. Three materials, including polyvinyl chloride, polyethylene terephthalate and polyimide, were used as substrates for fabricating ion-selective electrodes (ISEs) using stencil printing, screen-printing and laser engraving, respectively. We compared sensitivities of the ISEs to determine the best material for the fabrication process of the ISEs. The electrode surfaces were modified with various carbon nanomaterials including multi-walled carbon nanotubes, graphene, carbon black, and their mixed suspensions as the intermediate layer to enhance sensitivities of the electrodes. An automated 3D-printed robot was used for the drop-cast procedure during ISE fabrication to eliminate manual steps. The sensor array was optimized, and the detection limits were 10 -5  M, 10 -5  M and 10 -4  M for detection of K + , Na + and Ca 2+ ions, respectively. The sensor array integrated with a portable wireless potentiometer was used to detect K + , Na + and Ca 2+ in real urine and simulated sweat samples and results obtained were in agreement with ICP-OES with good recoveries. The developed sensing platform offers low-cost detection of electrolytes for point-of-care applications., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

A wireless-based detection utilizing an innovative electrochemical card (eCard) sensor controlled by a smartphone was developed for targeting Hepatitis B surface antigen (HBsAg). A simple label-free electrochemical platform allows a convenient operation for point-of-care diagnosis. A disposable screen-printed carbon electrode was modified straightforwardly layer-by-layer with chitosan followed by glutaraldehyde, allowing a simple but effective, reproducible, and stable method for covalently immobilizing antibodies. The modification and immobilization processes were verified by electrochemical impedance spectroscopy and cyclic voltammetry. The smartphone-based eCard sensor was used to quantify HBsAg by measuring the change in current response of the [Fe(CN) 6 ] 3-/4- redox couple before and after the presence of HBsAg. Under the optimal conditions, the linear calibration curve for HBsAg was found to be 10-100,000 IU/mL with a detection limit of 9.55 IU/mL. The HBsAg eCard sensor was successfully applied to detect 500 chronic HBV-infected serum samples with satisfactory results, demonstrating the excellent applicability of this system. The sensitivity and specificity of this sensing platform were found to be 97.75% and 93%, respectively. As illustrated, the proposed eCard immunosensor offered a rapid, sensitive, selective, and easy-to-use platform for healthcare providers to rapidly determine the infection status of HBV patients., (© 2023. The Author(s).)

3D printing technologies are an attractive for fabricating electrochemical sensors due to their ease of operation, freedom of design, fast prototyping, low waste, and low cost. We report the fabrication of a simple 3D-printed electrochemical sensing device for non-enzymatic detection of creatinine, an important indicator of renal function. To create the 3D-printed electrodes (3DE), carbon black/polylactic acid (CB/PLA) composite filament was used. The 3DE was activated using 0.5 M NaOH via amperometry prior to use to improve electrochemical performance. To give selectivity for creatinine, the activated 3DE was modified with a copper oxide nanoparticle-ionic liquid/reduced graphene oxide (CuO-IL/rGO) composite. The modified 3DE was characterized using microscopy and electrochemistry. Cyclic voltammetry and amperometry were used to evaluate sensor performance. The modified 3DE provided electrocatalytic activity towards creatinine without enzymes. Under optimal conditions, the modified 3DE directly coupled with a portable smartphone potentiostat exhibited the linear detection range of 0.5-35.0 mM, and the limit of detection was 37.3 μM, which is sufficient for detecting creatinine in human urine samples. Furthermore, the other physiological compounds present in human urine were not detected on the modified 3DE. Therefore, the modified 3DE could be a tool for effective creatinine screening in the urine., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

The COVID-19 pandemic has significantly increased the development of the development of point-of-care (POC) diagnostic tools because they can serve as useful tools for detecting and controlling spread of the disease. Most current methods require sophisticated laboratory instruments and specialists to provide reliable, cost-effective, specific, and sensitive POC testing for COVID-19 diagnosis. Here, a smartphone-assisted Sensit Smart potentiostat (PalmSens) was integrated with a paper-based electrochemical sensor to detect severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). A disposable paper-based device was fabricated, and the working electrode directly modified with a pyrrolidinyl peptide nucleic acid (acpcPNA) as the biological recognition element to capture the target complementary DNA (cDNA). In the presence of the target cDNA, hybridization with acpcPNA probe blocks the redox conversion of a redox reporter, leading to a decrease in electrochemical response correlating to SARS-CoV-2 concentration. Under optimal conditions, a linear range from 0.1 to 200 nM and a detection limit of 1.0 pM were obtained. The PNA-based electrochemical paper-based analytical device (PNA-based ePAD) offers high specificity toward SARS-CoV-2 N gene because of the highly selective PNA-DNA binding. The developed sensor was used for amplification-free SARS-CoV-2 detection in 10 nasopharyngeal swab samples (7 SARS-CoV-2 positive and 3 SARS-CoV-2 negative), giving a 100% agreement result with RT-PCR., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Nucleic acid biosensors for point-of-care (POC) diagnostic applications are highly desirable. The ability to detect DNA and RNA in a simple, rapid, affordable and portable format leads to a range of important applications for early screening in the field of disease monitoring and management. Herein, we report the development of an isothermal, label-free electrochemical biosensor that was designed on the basis of target-driven MNAzyme cleavage activity. Hybridization with HPV mRNA, a model nucleic acid target, activated MNAzyme and initiated the cleavage of immobilized hairpin substrates, leading to changes in the electrochemical signal. Under optimal conditions, a detection limit of 2.6 pM was obtained with an incubation time of 60 min. Furthermore, an artificial chaperone-enhanced MNAzyme (ACEzyme) system was integrated to an electrochemical biosensor for the first time. The analytical performance of the biosensor was enhanced, and the detection time was significantly reduced by the addition of PLL-g-Dex, which exhibits nucleic acid chaperone-like activity. A detection limit of 0.88 pM was obtained with a threefold decrease in incubation time without prior amplification. The proposed biosensing platform shows the advantages of simple fabrication and operation, good selectivity in the presence of single-base mismatch, and excellent versatility in a complex mixture of total RNA. We believe that this isothermal, label-free, and protein-free nucleic acid analysis platform could provide foundations for the further development of a universal nucleic acid biosensing platform for clinical application., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

The current approaches of diagnostic platforms for detecting SARS-CoV-2 infections mostly relied on adapting the existing technology. In this work, a simple and low-cost electrochemical sensing platform for detecting SAR-CoV-2 antigen was established. The proposed sensor combined the innovative disposable paper-based immunosensor and cost-effective plant-based anti-SARS-CoV-2 monoclonal antibody CR3022, expressed in Nicotiana benthamiana. The cellulose nanocrystal was modified on screen-printed graphene electrode to provide the abundant COOH functional groups on electrode surface, leading to the high ability for antibody immobilization. The quantification of the presence receptor binding domain (RBD) spike protein of SARS-CoV-2 was performed using differential pulse voltammetry by monitoring the changing current of [Fe(CN) 6 ] 3-/4- redox solution. The current change of [Fe(CN) 6 ] 3-/4- before and after the presence of target RBD could be clearly distinguished, providing a linear relationship with RBD concentration in the range from 0.1 pg/mL to 500 ng/mL with the minimum limit of detection of 2.0 fg/mL. The proposed platform was successfully applied to detect RBD in nasopharyngeal swab samples with satisfactory results. Furthermore, the paper-based immunosensor was extended to quantify the RBD level in spiked saliva samples, demonstrating the broadly applicability of this system. This electrochemical paper-based immunosensor has the potential to be employed as a point-of-care testing for COVID-19 diagnosis., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

In this work, an automated flow injection analysis (FIA) connected to a boron-doped diamond electrode (BDDE) was originally developed for the analysis of methimazole in pharmaceutical preparations. At a modification-free BDDE, methimazole was easilly oxidized. For the analysis of the mechanisms occurring at the electrode surface, cyclic voltammetry was employed to evaluate the impact of fundamental experimental parameters, such as pH and scan rate, on the BDDE response. For the quantitative detection, the FIA amperometric approach was constructed and used as a fast and sensitive method. The suggested approach provided a broad linear range of 0.5-50 μmol/L and a low detection limit of 10 nmol/L (signal-to-noise ratio = 3). Furthermore, the BDDE was successfully utilized to quantify methimazole in genuine samples from a variety of medicines, and its performance remained steady after more than 50 tests. The findings of amperometric measurements exhibit excellent repeatability, with relative standard deviations of less than 3.9 and 4.7 % for intra-day and inter-day, respectively. The findings indicated that, compared with traditional approaches, the suggested method has the following advantages: quick analysis time, simplicity, highly sensitive output, and no need for complicated operational processes., Competing Interests: Conflicts of interest: All the authors declare no conflict of interest., (Copyright © 2023 by the authors.)

Cardiac troponin I (cTnI) is a specific cardiac biomarker for diagnosis of acute myocardial infarction (AMI). A sensitive and simple point-of-care test (POCT) is still required for early detection of AMI. To address this need, we developed a dip strip assay based on sandwich immunoassay coupled with a silver enhancement system. Pre-incubation and silver enhancement were introduced to the dip strip to increase sensitivity. Due to the catalytic reaction of the silver enhancement solution, the red color of AuNPs changed to dark brown as silver ions precipitated and enlarged the AuNPs. The obtained results were easily seen by the naked eye. For quantitative analysis, the color intensity of the results was analyzed using a smartphone with RGB color picker application. The effects of operating parameters (volume of AuNP-Ab conjugate, volume of sample, incubation time, and analysis time) were investigated and optimized. Under optimal conditions, the limit of detection (LOD) by the naked eye was 0.5 ng/mL. The LOD with silver enhancement was 50-fold lower than without. For quantitative analysis using the smartphone, linearity of detection was observed through the range of 0.5-50 ng/mL (R 2  = 0.9952) and the LOD was 0.12 ng/mL. The developed method was successfully applied to detection of cTnI in serum samples, achieving analytical recoveries and %RSD in the ranges of 96.10-119.17% and 2.91-5.13%, respectively. Additionally, this developed assay was not cross reactive with the potentially interfering serum proteins. These results showed the great potential of this dip strip assay as an alternative POCT for detection of serum cTnI., (© 2022. The Author(s).)

A facile colorimetric method based on a typical redox reaction was first developed for the determination of salbutamol (SAL) using a low-cost and portable transparency-based analytical device (TAD). The TAD was simply fabricated by wax-printing onto a transparent polymer-based substrate to create the hydrophobic barriers and the colorimetric reaction zones where the color changes could be easily observed with the naked eye. Potassium permanganate (KMnO 4 ), a common oxidizing agent, was deliberately used as a colorimetric reagent for SAL. Once SAL reacted with KMnO 4 in the acidified system, it could undergo oxidation and the color of KMnO 4 subsequently changed from light pink to orange. The color change corresponding to the SAL concentration could be clearly observed at the TAD sensor. In addition, the reaction color could be recorded using a digital camera and then analyzed by ImageJ for quantitative analysis. Under the optimized conditions, the developed method together with the TAD sensor exhibited high efficiency for SAL determination with linearity ranging from 0.5 to 40 mg·L -1 and a limit of detection (LOD) of 0.05 mg·L -1 . •This proposed TAD-based colorimetric method using permanganate as color reagent showed excellent performance in SAL detection with good accuracy and high precision., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2022 The Author(s).)

Leptospirosis is one of the most life-threatening tropical diseases caused by pathogenic Leptospira . To date, a diagnostic device that offers rapid and sensitive detection of leptospires has been still in demand for proper treatment to reduce the mortality rate. Herein, we create a resistance-based lateral flow immunosensor diagnosis device (R-LFI) that integrates near-field communication (NFC) with a portable smartphone for leptospiral detection in clinical samples. A specific monoclonal antibody against the pathogen was coated on a nitrocellulose membrane (NCM) where the test line was collocated. Two electrodes with a sandwich-like configuration were installed employing a conductive double-sided adhesive tape and connected with a NFC smartphone-based detection system. A half-sandwich immunocomplex formation induced high proton conduction, resulting in a considerable decrement in resistive response. The performance of the R-LFI sensor was evaluated using recombinant LipL32 (rLipL32), Leptospira interrogans , and clinical samples. The R-LFI device exhibited linear responses toward rLipL32 protein in phosphate buffer and L. interrogans -spiked healthy human serum samples within the concentration ranging from 1 to 1000 ng mL -1 (limit of detection (LOD): 0.29 ng mL -1 ) and from 10 4 to 10 6 cell mL -1 (LOD: 4.89 × 10 3 cell mL -1 ), respectively. Our R-LFI sensor successfully detected L. interrogans -positive clinical samples as confirmed by polymerase chain reaction (PCR). This platform offers high specificity, selectivity, simplicity, miniscule sample volume, and no labeling element requirement. These desirable features make it particularly suitable for countries where medical facilities and resources are limited.

A new detection strategy was developed to improve the sensitivity of a lateral flow immunoassay platform utilizing a delayed hydrophobic barrier fabricated with trimethylsilyl cellulose (TMSC). The SARS-CoV-2 spike receptor-binding domain (SARS-CoV-2 SP RBD) antigen was chosen as a model analyte to demonstrate the superior detectability of this scheme. The novel device consists of 2 separate layers, so-called delayed lateral flow immunoassay (d-LFIA). The upper layer is intended for the analyte or sample flow path, where the test solution flows freely straight to the detection zone to bind with the primary antibody. The lower layer, located just underneath, is designed for the SARS-CoV-2 spike receptor-binding domain-conjugated gold nanoparticles (SARS-CoV-2 SP RBD-AuNPs) used for producing a colorimetric signal. This layer is fabricated with a TMSC barrier to time-delay the movement of SARS-CoV-2 SP RBD-AuNPs, thus allowing the antigen to bind with the primary antibody more efficiently. This platform exhibited a 2.6-fold enhancement in the sensitivity and 9.1-fold improvement in the limit of detection (LOD) as compared with the conventional LFIA. In addition, this d-LFIA device was satisfactorily applied to accurate screening of COVID-19 patients., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

Pt nanoparticles deposited on single-walled carbon nanotubes (PtSWCNTs), synthesized via the deposition precipitation (DP) method, were introduced as a substrate for immobilizing antibodies on an electrode surface and then enhancing the electrochemical sensitivity. A PtSWCNT-modified paper-based screen-printed graphene electrode was successfully developed to diagnose hepatitis C virus (HCV) infection. The hepatitis C virus core antigen (HCV-cAg) level was determined by differential pulse voltammetry (DPV) using [Fe(CN) 6 ] 3-/4- as a redox solution. In the presence of HCV-cAg, the DPV current response decreased with increasing HCV-cAg concentration. Under the optimal conditions, the change in current response provides a good linear correlation with the logarithm of HCV-cAg concentration in the range 0.05 to 1000 pg mL -1 (RSD < 5%), and the limit of detection was 0.015 pg mL -1 (or 0.71 fmol L -1 ). Furthermore, the proposed immunosensor has been utilized to quantify HCV-cAg in human serum samples with reliable results compared with standard immunoassays (% relative error < 10%). This sensor offers a simple, sensitive, selective, disposable, and inexpensive means for determination of HCV-cAg in human serum samples. The paper-based label-free immunosensor is versatile and feasible for clinical diagnosis., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

Early determination of blood lactate levels may accelerate the detection of sepsis, one of the most time-sensitive illnesses. We developed and validated a portable blood lactate detection kit for clinical screening that can measure early bedside lactate levels in intensive care unit (ICU) patients suspected of having sepsis. A TiO 2 sol-G nanocomposite was prepared and coated on a screen-printed carbon electrode (SPCE) integrated with non-immobilized lactate oxidase (LOx) to produce a novel lactate biosensor with high sensitivity and high storage stability for human blood lactate measurement. The detection kit was based on an electrochemical technique and showed a wide linear range of 1-20 mM ( R 2 = 0.9937) with a low detection limit of 0.2 mM for lactate detection. This allowed for differentiating patient groups who may have sepsis using a cut-off level of 4 mM. The device was successfully implemented for blood lactate determination in critical patients, showing an accuracy range from 75% to 112%. This device provided high-precision and rapid quantitative information validated using a blood gas analyzer. Our detection kit might help to reduce the morbidity and mortality rates in severe sepsis and septic shock patients in community hospitals.

Among lateral flow immunoassay (LFIA) platforms, enzyme-based LFIAs provide signal amplification to improve sensitivity. However, most enzyme-based LFIAs require multiple timed steps, complicating their utility in point-of-care testing (POCT). Here, we report a microfluidic interface for LFIAs that automates sample, buffer, and reagent addition, greatly simplifying operation while achieving the high analytical stringency associated with more complex assays. The microfluidic interface also maintains the low cost and small footprint of standard LFIAs. The platform is fabricated from a combination of polyester film, double-sided adhesive tape, and nitrocellulose, and fits in the palm of your hand. All reagents are dried on the nitrocellulose to facilitate sequential reagent delivery, and the sample is used as the wash buffer to minimize steps. After the sample addition, a user simply waits 15 min for a colorimetric result. This manuscript discusses the development and optimization of the channel geometry to achieve a simple step enzyme amplified immunoassay. As a proof-of-concept target, we selected lipoarabinomannan (LAM), a WHO identified urinary biomarker of active tuberculosis, to demonstrate the device feasibility and reliability. The results revealed that the device successfully detected LAM in phosphate buffer (PBS) as well as spiked urine samples within 15 min after sample loading. The minimum concentration of color change was achieved at 25 ng mL -1 .

Thread-based electrochemical immunosensor is fabricated for non-invasive detection of cortisol in sweat by immobilization of anti-cortisol on L-cys/AuNPs/MXene modified conductive thread electrode. MXene and AuNPs increase the surface area of conductive thread electrode and facilitate anti-cortisol immobilization leading to enhanced sensor sensitivity. Anti-cortisol is immobilized on L-cys/AuNPs/MXene modified electrode by using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide and N-hydroxysulfosuccinimide coupling agents. The electrochemical detection of cortisol is based on the decrease of oxidation current towards the antigen-antibody binding interaction owing to blocking of electron transfer process by cortisol. Under the optimal conditions, this immunosensor offers high sensitivity, a wide linearity of 5-180 ng mL -1 and a detection limit of 0.54 ng mL -1 with negligible effect from interferences. Furthermore, this immunosensor provides high reproducibility and long-term storage stability (≥6 weeks). Ultimately, this system is successfully applied for the detection of cortisol in artificial sweat with satisfactory results. Hence, this platform might be suitable to apply as a wearable electrochemical sensor for sweat cortisol by integrating on a wristband., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Screen-printed graphene electrodes (SPGEs) have become a potential option in electrochemical applications because of their outstanding properties and disposable approach to miniaturize the electrodes for onsite analysis. Herein, the detection of para -hydroxybenzoic acid (PHBA) in cosmetics using the anodized SPGE has been pioneered and reported. The simple anodization of the SPGE surface was operated by anodic pretreatment at a constant potential on SPGE. The surface morphologies and electrochemical behaviors of anodized SPGEs in different anodization electrolytes were examined. Using anodized SPGE in a phosphate-buffered solution, a nontoxic solution, the sensitivity of PHBA detection was significantly improved compared with pristine SPGE owing to the increase of the polar oxygen-containing functional group during the anodization. The anodized SPGE could detect a PHBA down to 0.073 μmol/L. Finally, the developed anodized SPGE presented high ability and feasibility for PHBA detection in cosmetics. Furthermore, a facile electrode preparation step with a nontoxic solution can present high reproducibility and compatibility with a portable potentiostat for onsite PHBA detection during manufacturing., Competing Interests: The authors declare no competing financial interest., (© 2022 The Authors. Published by American Chemical Society.)

The electrochemical determination of five heavy metals is demonstrated using a wireless and card-sized potentiostat coupled with a smartphone through near-field communication (NFC) technology. A smartphone application was customized to command the NFC potentiostat, collect real-time signals, process the data, and ultimately display the quantities of the selected elements. The screen-printed graphene electrode (SPGE) was simply fabricated and modified using different nanomaterials for each heavy metal. Using differential pulse voltammetry (DPV) mode on the smartphone, the signal peaks were presented at + 10 mV for As(III), + 350 mV for Cr(VI), 0 mV for Hg(II), - 900 mV for Cd(II), and - 680 mV vs. Ag/AgCl for Pb(II). The linear ranges were 25-500, 250-25,000, 100-1,500, 25-750, 25-750 ng mL -1 with detection limits of 3.0, 40, 16, 2.0, and 0.95 ng mL -1 for As(III), Cr(VI), Hg(II), Cd(II), and Pb(II), respectively. The reproducibility in terms of relative standard deviation was less than 8.8% (n = 5 devices) of the developed SPGE coupled with the NFC potentiostat. Various samples for different applications (e.g., food safety and environmental monitoring) were analyzed and quantified using the proposed sensors. The results from this sensor indicate that there is no significant difference (95% confidence level) compared with those obtained from the traditional ICP-OES method, while the recoveries were found in the acceptable range of 80-111%. Hence, it can be deduced that this recent advanced technology of the NFC potentiostat developed for heavy metal analysis offers a highly sensitive and selective detection, yet the sensor remains compact, low-cost, and readily accessible to end-users., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

LipL32 is an outer membrane protein present only on pathogenic Leptospira species, which is the causative agent of leptospirosis. Leptospirosis symptoms are often misdiagnosed with other febrile illnesses as the clinical manifestations are non-specific. Therefore, an accurate diagnostic tool for leptospirosis is indeed critical for proper and prompt treatment. Typical diagnosis via serological assays is generally performed to assess the antibodies produced against Leptospira . However, their delayed antibody response and complicated procedure undoubtedly limit the practical utilization especially in a primary care setting. Here, we demonstrate for the first time an early-stage detection of LipL32 by an integrated lateral-flow immunoassay with an electrochemical readout (eLFIA). A ferrocene trace tag was monitored via differential pulse voltammetry operated on a smartphone-based device, thus allowing for on-field testing. A superior performance in terms of the lowest detectable limit of detection of 8.53 pg/mL and broad linear dynamic range (5 orders of magnitude) among other sensors available thus far was established. Additionally, the developed test strip provided a straightforward yet sensitive approach for diagnosis of leptospirosis using the collected human sera from patients, in which the results were comparable to the real-time polymerase chain reaction technique.

Coinfection of HIV/HCV is a significant public health issue globally, as it increases the risk of liver cancer in co-infected individuals. The point-of-care testing (POCT) device for HIV/HCV DNA detection is promptly needed for diagnosis and monitoring of the disease progression. Here, the alternating-current electroluminescence (ACEL) technique is proposed as a sensitive POCT sensing platform for HIV/HCV cDNA detection. A conductance-based light emission modulated by the hybridization between a pyrrolidinyl PNA probe and the DNA target enabled the DNA detection in a label-free format. Enhanced electroluminescence was observed in the presence of the target DNA due to the increased proton conductivity. Under the optimal conditions, the linearity range from 1 nM to 1 μM was achieved for HIV and HCV cDNA with LODs of 1.86 pM (HIV cDNA) and 1.96 pM (HCV cDNA). The spiked HIV/HCV cDNA in healthy human serum was successfully detected, demonstrating the feasibility of the developed device for the detection of cDNA in real biological samples. Additionally, simultaneous HIV/HCV cDNA detection on a single ACEL device employing a 2x2-array detection zone design. The cross-reactivity with other viral DNA was shown to be minimal due to the high specificity of the PNA probes used. Finally, the negative and positive samples from the patient's serum were tested and the results were in 100% agreement with the commercial kit based-on real-time PCR method, thus illustrating the high sensitivity and specificity of the developed sensor., (Copyright © 2021. Published by Elsevier B.V.)

Background: The demand for point-of-care testing (POCT) devices has rapidly grown since they offer immediate test results with ease of use, makingthem suitable for home self-testing patients and caretakers. However, the POCT development has faced the challenges of increased cost and limited resources. Therefore, the paper substrate as a low-cost material has been employed to develop a cost-effective POCT device, known as "Microfluidic paper-based analytical devices (μPADs)". This device is gaining attention as a promising tool for medicinal diagnostic applications owing to its unique features of simple fabrication, low cost, enabling manipulation flow (capillarydriven flow), the ability to store reagents, and accommodating multistep assay requirements., Objective: This review comprehensively examines the fabrication methods and device designs (2D/3D configuration) and their advantages and disadvantages, focusing on updated μPADs applications for motif identification., Methods: The evolution of paper-based devices, starting from the traditional devices of dipstick and lateral flow assay (LFA) with μPADs, has been described. Patterned structure fabrication of each technique has been compared among the equipment used, benefits, and drawbacks. Microfluidic device designs, including 2D and 3D configurations, have been introduced as well as their modifications. Various designs of μPADs have been integrated with many powerful detection methods such as colorimetry, electrochemistry, fluorescence, chemiluminescence, electrochemiluminescence, and SER-based sensors for medicinal diagnosis applications., Conclusion: The μPADs potential to deal with commercialization in terms of the state-of-the-art of μPADs in medicinal diagnosis has been discussed. A great prototype, which is currently in a reallife application breakthrough, has been updated., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

In the last decade, due to the global increase in diseases, drugs for biomedical applications have increased dramatically. Therefore, there is an urgent need for analytical tools to monitor, treat, investigate, and control drug compounds in diverse matrices. The new and challenging task has been looking for simple, low-cost, rapid, and portable analytical platforms. The development of microfluidic paper-based analytical devices (μPADs) has garnered immense attention in many analytical applications due to the benefit of cellulose structure. It can be functionalized and serves as an ideal channel and scaffold for the transportation and immobilization of various substances. Microfluidic technology has been considered an effective tool in pharmaceutical analysis that facilitates the quantitative measurement of several parameters on cells or other biological systems. The μPADs represent unique advantages over conventional microfluidics, such as the self-pumping capability. They have low material costs, are easy to fabricate, and do not require external power sources. This review gives an overview of the current designs in this decade for μPADs and their respective application in pharmaceutical analysis. These include device designs, choice of paper material, and fabrication techniques with their advantages and drawbacks. In addition, the strategies for improving analytical performance in terms of simplicity, high sensitivity, and selectivity are highlighted, followed by the application of μPADs design for the detection of drug compounds for various purposes. Moreover, we present recent advances involving μPAD technologies in the field of pharmaceutical applications. Finally, we discussed the challenges and potential of μPADs for the transition from laboratory to commercialization., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

Electrochemical paper-based analytical devices (ePADs) are useful analytical devices that serve as point-of-care testing (POCT) devices for various clinical biomarkers in view of their simplicity, portability, and low-cost format. However, multistep reagent manipulation usually restricts the performance of the device for end users. Herein, we developed a sequential ePAD for sequential immunosensing fluid delivery by integrating dual flow behaviors (fast-flow/delayed) within a single paper platform for the simultaneous detection of hepatitis B surface antigen (HBsAg) and hepatitis C core antigen (HCVcAg). In the present work, a fast-flow channel was used for the automated washing of unbound antigens, while a delayed channel was created to store a redox reagent for further electrochemical analysis with a single buffer loading (the analysis time can be completed within 500 s). Hence, the undesirable complex procedure of multi-step reagent manipulation is scarcely needed by the user. The detection limit of the proposed ePAD was as low as 18.2 pg mL -1 for HBsAg and 1.19 pg mL -1 for HCVcAg. In addition, this proposed ePAD was also proven to be effective in real clinical sera from patients to verify its biological applicability. The ePAD sensor shows high promise as an easy-to-use, portable, and extendable sensor for other multiplex biological assays., (Copyright © 2021 Elsevier B.V. All rights reserved.)