Nanomaterial-based Electrochemical Sensors for Multiplex Medicinal Applications.

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.

Sequential Flow Controllable Microfluidic Device for G-Quadruplex DNAzyme-Based Electrochemical Detection of SARS-CoV-2 Using a Pyrrolidinyl Peptide Nucleic Acid.

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 H2O2-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.

Electrochemical Capillary Driven Immunoassay for Detection of SARS-CoV-2.

The COVID-19 pandemic focused attention on a pressing need for fast, accurate, and low-cost diagnostic tests. This work presents an electrochemical capillary driven immunoassay (eCaDI) developed to detect SARS-CoV-2 nucleocapsid (N) protein. The low-cost flow device is made of polyethylene terephthalate (PET) and adhesive films. Upon addition of a sample, reagents and washes are sequentially delivered to an integrated screen-printed carbon electrode for detection, thus automating a full sandwich immunoassay with a single end-user step. The modified electrodes are sensitive and selective for SARS-CoV-2 N protein and stable for over 7 weeks. The eCaDI was tested with influenza A and Sindbis virus and proved to be selective. The eCaDI was also successfully applied to detect nine different SARS-CoV-2 variants, including Omicron.

Recent developments in microfluidic paper-based analytical devices for pharmaceutical analysis.

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.

An application of miniaturized electrochemical sensing for determination of arsenic in herbal medicines.

This study aimed to create a miniaturized electrochemical platform for detecting As(III) contamination in herbal medicines. To reduce the operational steps of modification and determination, only a single drop of mixed standard Au(III) and sample solution is proposed to perform the electrochemical measurements using a screen-printed graphene electrode (SPGE). Square wave anodic stripping voltammetry was employed to integrate the simultaneous modification and determination processes. To perform the measurement, As(III) and Au(III) migrate to the SPGE surface while the reduction potential is held at -0.5 V, forming an Au-As intermetallic alloy. Then, As is stripped off for the electrochemical determination of As(III). The total assay time is less than 3 min. Under suitable conditions, the electrochemical sensing system can detect As(III) at concentrations ranging from 0.1 to 3.0 ppm, with a limit of quantification and limit of detection of 0.1 and 0.03 ppm, respectively. The applicability and accuracy of the proposed sensor were verified by determining As(III) in herbal medicinal samples, and they were found to be in line with the standard method (ICP-OES). The benefits of simple operation, rapid detection, portability, and low cost (<1 USD) make this a more powerful tool for routine monitoring and on-site analysis applications.

Electrochemical Immunoassay for the Detection of SARS-CoV-2 Nucleocapsid Protein in Nasopharyngeal Samples.

Point-of-care (POC) methods currently available for detecting SARS-CoV-2 infections still lack accuracy. Here, we report the development of a highly sensitive electrochemical immunoassay capable of quantitatively detecting the presence of the SARS-CoV-2 virus in patient nasopharyngeal samples using stencil-printed carbon electrodes (SPCEs) functionalized with capture antibodies targeting the SARS-CoV-2 nucleocapsid protein (N protein). Samples are added to the electrode surface, followed by horseradish peroxidase (HRP)-conjugated detection antibodies also targeting the SARS-CoV-2 N protein. The concentration of the virus in samples is quantified using chronoamperometry in the presence of 3,3'5,5'-tetramethylbenzidine. Limits of detection equivalent to less than 50 plaque forming units/mL (PFU/mL) were determined with virus sample volumes of 20 µL. No cross-reactivity was detected with the influenza virus and other coronavirus N proteins. Patient nasopharyngeal samples were tested as part of a proof-of-concept clinical study where samples were also tested using the gold-standard real-time quantitative polymerase chain reaction (RT-qPCR) method. Preliminary results from a data set of 22 samples demonstrated a clinical specificity of 100% (n = 9 negative samples according to RT-qPCR) and a clinical sensitivity of 70% for samples with RT-PCR cycle threshold (Ct) values under 30 (n = 10) and 100% for samples with Ct values under 25 (n = 5), which complies with the World Health Organization (WHO) criteria for POC COVID-19 diagnostic tests. Our functionalized SPCEs were also validated against standard plaque assays, and very good agreement was found between both methods (R2 = 0.9993, n = 6), suggesting that our assay could be used to assess patient infectivity. The assay currently takes 70 min from sampling to results.

A novel l-cysteine sensor using in-situ electropolymerization of l-cysteine: Potential to simple and selective detection.

This work presents an all-in-one origami paper-based electrochemical platform for simple and inexpensive l-cysteine (Cys) detection using Cys as a monomer for modifying electrode surfaces. The proposed method combines the steps of electropolymerization and detection into a single device to offer a highly convenient method for the end-user. In comparison, the sensitivity toward Cys detection is a significantly increased using this modified electrode. The developed device provided a linear concentration range of 10-800 µM with a limit of detection of 5.5 µM. For application, the device was successfully applied to detect Cys in different food products such as wheat flour, bread, and cake with satisfactory results, yielding excellent intra-day and inter-day relative standard deviations (1.5-4.9%) and recoveries (84.2-110.8%). This discovery is important from the viewpoint of the development of Cys detection in other applications in the future.

Electrochemical Capillary-Flow Immunoassay for Detecting Anti-SARS-CoV-2 Nucleocapsid Protein Antibodies at the Point of Care.

Rapid and inexpensive serological tests for SARS-CoV-2 antibodies are needed to conduct population-level seroprevalence surveillance studies and can improve diagnostic reliability when used in combination with viral tests. Here, we report a novel low-cost electrochemical capillary-flow device to quantify IgG antibodies targeting SARS-CoV-2 nucleocapsid proteins (anti-N antibody) down to 5 ng/mL in low-volume (10 µL) human whole blood samples in under 20 min. No sample preparation is needed as the device integrates a blood-filtration membrane for on-board plasma extraction. The device is made of stacked layers of a hydrophilic polyester and double-sided adhesive films, which create a passive microfluidic circuit that automates the steps of an enzyme-linked immunosorbent assay (ELISA). The sample and reagents are sequentially delivered to a nitrocellulose membrane that is modified with a recombinant SARS-CoV-2 nucleocapsid protein. When present in the sample, anti-N antibodies are captured on the nitrocellulose membrane and detected via chronoamperometry performed on a screen-printed carbon electrode. As a result of this quantitative electrochemical readout, no result interpretation is required, making the device ideal for point-of-care (POC) use by non-trained users. Moreover, we show that the device can be coupled to a near-field communication potentiostat operated from a smartphone, confirming its true POC potential. The novelty of this work resides in the integration of sensitive electrochemical detection with capillary-flow immunoassay, providing accuracy at the point of care. This novel electrochemical capillary-flow device has the potential to aid the diagnosis of infectious diseases at the point of care.