Electrochemical Immunoassay Research Articles

Designable Electrochemiluminescence Patterning for Renewable and Enhanced Bioimaging.

Electrochemical imaging enables an in-depth analysis of the interface heterogeneity and reaction kinetics of single entities. However, electrode passivation during electrochemical reactions decreases the active sites and harms the long-term stability. Here, we introduce a method using laser-induced photothermal effects to restore the electrochemical activity, which is particularly displayed as enhanced micrometric patterns in electrochemiluminescence (ECL) microscopy. By co-localization characterization and X-ray photoelectron spectroscopy, the mechanism of active site regeneration is validated as the removal of the oxide film for restoring the local surface ECL reactivity under laser irradiation. The surface-confined and voltage-dependent features of ECL allows for easy pattern erasure and rewriting, and it shows good reversibility and anti-counterfeiting potential. This approach overcomes the passivation processes, evidently improves the image quality of single biological entities including Shewanella bacteria and cells, and makes the subtle contour structures more distinct. The renewable electrode interface also enhances the ECL signal of model bead-based bioassays. This approach not only showcases precise control in fabricating micron patterns but also holds promise for enhancing the sensitivity in electrochemical immunoassays and bioimaging.

Designable Electrochemiluminescence Patterning for Renewable and Enhanced Bioimaging

Electrochemical imaging enables an in‐depth analysis of the interface heterogeneity and reaction kinetics of single entities. However, electrode passivation during electrochemical reactions decreases the active sites and harms the long‐term stability. Here, we introduce a method using laser‐induced photothermal effects to restore the electrochemical activity, which is particularly displayed as enhanced micrometric patterns in electrochemiluminescence (ECL) microscopy. By co‐localization characterization and X‐ray photoelectron spectroscopy, the mechanism of active site regeneration is validated as the removal of the oxide film for restoring the local surface ECL reactivity under laser irradiation. The surface‐confined and voltage‐dependent features of ECL allows for easy pattern erasure and rewriting, and it shows good reversibility and anti‐counterfeiting potential. This approach overcomes the passivation processes, evidently improves the image quality of single biological entities including Shewanella bacteria and cells, and makes the subtle contour structures more distinct. The renewable electrode interface also enhances the ECL signal of model bead‐based bioassays. This approach not only showcases precise control in fabricating micron patterns but also holds promise for enhancing the sensitivity in electrochemical immunoassays and bioimaging.

Dual on-the-move electrochemical immunoassays for the simultaneous determination of amyloid-β (1−42) and Tau in Alzheimer's patient samples

Here, we report catalytic micromotors (MM)-based electrochemical immunoassays for the on-the-move dual and simultaneous determination of Amyloid-β (Aβ-42) and Tau protein (Tau) (MMAβ-42-MMTau) as relevant Alzheimer’s disease biomarkers in brain tissue, cerebrospinal fluid, and plasma diagnosed samples. Combining the binding capacity of the antibody's functionalized polypyrrole (PPy) layer of MM with the self-propulsion from the PtNPs layer thanks to the decomposition of hydrogen peroxide, the approach yielded excellent detection limits (LODAβ-42=0.04 ng/mL, LODTau= 0.4 pg/mL) using low sample volumes (30 µL) and short analysis times (15 min) to detect both biomarkers. Quantitative analysis by MMAβ-42-MMTau was carried out without any clinical sample dilution (linear ranges are between 0.1 and 5 ng/mL for Aβ-42 and from 1 to 106 pg/mL in the case of Tau), highlighting the versatility of the approach to quantify Aβ-42 and Tau levels at different dynamic ranges. MMAβ-42-MMTau showed superior analytical capabilities to the single molecule counting technology (SMCx) during quantitative analysis in all sample classes tested, reporting a difference in quantitative levels for both biomarkers between healthy and diseased individuals and an increase in the levels with disease progression, except in plasma samples where no relationship between biomarker levels and disease progression was found.

Investigation and Demonstration for immunoassay technology based on impedance Amplifies of magnetic and dielectric microbeads

BackgroundBead-based immunoassays typically require researchers to perform the analysis in a laboratory setting, involving complex processing procedures, most of the time, constant potential or current instruments are used to measure the bio-impedance change. However, almost all biological materials are decorated on the electrode surface to obtain the maximum sensitivity, resulting in difficulty to clean, separate, and other applications of fixed bead-based immunoassay. ResultWe present an innovative unfixed bead-based immunoassay technology that utilizes electrical impedance characteristics, offering the advantages of being rapid, portable, miniaturized, and cost-effective. This technique which is known as Impedance-enhanced electrochemical immunoassay (IE-ECIA) employs antibody-coated Magnetic beads (MBs) and Dielectric beads (DBs) linked to antigens for quantitative with miniaturization electronic devices, which has been successfully applied in an immune impedance sensor for detecting the Dengue virus (DENV), which is an affordable test with a low-cost microfluidic impedance biosensor combined with immune magnetic and dielectric micro/nanobeads for antigen detection. HighlightThe proposed immunoassay technology introduces a novel microfluidic platform incorporating magnetic and dielectric beads alongside with an electrochemical impedance biosensor, therefore facilitating continuous-fluid quantitative analysis. This innovative and unconventional approach leverages free-motion multiplex beads housing magnetic and dielectric bead-bound antigens within a simple linear microchannel of microfluidic chip architecture, making the high-through measurements more efficient, enhanced Electrochemical immunoassay (ECIA) signals of Bio-on-magnetic-beads (BOMB). This advancement holds significant potential for enhancing diagnostic capabilities for emerging infectious diseases within clinical environments, thereby bolstering rapid detection capabilities and aiding in the advancement of novel pharmaceuticals, benefiting all the stakeholders.

Electrografted Laser-Induced Graphene: Direct Detection of Neurodegenerative Disease Biomarker in Cerebrospinal Fluid.

There are more than 50 neurodegenerative disorders, and amyotrophic lateral sclerosis (ALS) is one of the most common disorders that poses diagnostic and treatment challenges. The poly glycine-proline (polyGP) dipeptide repeat is a toxic protein that has been recognized as a pharmacodynamic biomarker of C9orf72-associated (c9+) ALS, a subtype of ALS that originates from genetic mutation. Early detection of polyGP will help healthcare providers start timely gene therapy. Herein, we developed a label-free electrochemical immunoassay for the simple detection of polyGP in unprocessed cerebrospinal fluid (CSF) samples collected from ALS patients in the National ALS Biorepository. For the first time, an electrografted laser-induced graphene (E-LIG) electrode system was employed in a sandwich format to detect polyGP using a label-free electrochemical impedance technique. The results show that the E-LIG-modified surface exhibited high sensitivity and selectivity in buffer and CSF media with limit of detection values of 0.19 and 0.27 ng/mL, respectively. The precision of the calibration model was better in CSF than in the buffer. The E-LIG immunosensor can easily select polyGP targets in the presence of other dipeptide proteins translated from the c9 gene. Further study with CSF samples from ALS patients demonstrated that the label-free E-LIG-based immunosensor not only quantified polyGP in the complex CSF matrix but also distinguished between c9+ and non-c9- ALS patients.

Overview of the Design and Application of Dual-Signal Immunoassays

Immunoassays have been widely used for the determination of various analytes in the fields of disease diagnosis, food safety, and environmental monitoring. Dual-signal immunoassays are now advanced and integrated detection technologies with excellent self-correction and self-validation capabilities. In this work, we summarize the recent advances in the development of optical and electrochemical dual-signal immunoassays, including colorimetric, fluorescence, surface-enhanced Raman spectroscopy (SERS), electrochemical, electrochemiluminescence, and photoelectrochemical methods. This review particularly emphasizes the working principle of diverse dual-signal immunoassays and the utilization of dual-functional molecules and nanomaterials. It also outlines the challenges and prospects of future research on dual-signal immunoassays.

Determination of deoxynivalenol (DON) by a label-free electrochemical immunosensor based on Ni[sbnd]Fe PBA nanozymes

Deoxynivalenol (DON) contamination in food products significantly threatens human health, necessitating a reliable and sensitive detection method. This study aims to develop a simple, low-cost, and effective electrochemical immunoassay method for detecting DON based on the nickel‑iron bimetallic Prussian blue analog (NiFe PBA). The NiFe PBA nanozymes with high peroxidase-like activity were synthesized using an environmentally friendly chemical precipitation method. In the presence of hydrogen peroxide (H2O2), the current change of thionine oxidation initiated by NiFe PBA nanozymes can be exploited to diagnose DON. Under optimal conditions, the proposed method achieved quantitative detection of DON in the range of 10–107 pg mL−1 with a detection limit of 4.5 pg mL−1 (S/N = 3), demonstrating excellent selectivity, reproducibility, and stability. In addition, the DON immunosensor provides satisfactory results for the detection in real samples, demonstrating the feasibility of the proposed sensor in detecting of DON in such products.

Effect of nanoarray density on enhanced electron transfer efficiency and analytical sensitivity for electrochemical immunosensors

The fabrication of ordered nanoarray electrode (NAE) using UV imprinting and their application as electrochemical (EC) immunosensor is described in this study. Especially, the influence of the array density factors on the performance of NAE was characterized electrochemically and compared with flat-electrode. Low-density (hole: 200 nm, hole space = 600 nm), medium-density (hole: 200 nm, hole space = 400 nm), and high-density NAE (hole: 200 nm, hole space = 200 nm) which have the same active area were fabricated and their redox cycling was compared with empirical results. We observed that the high-density is the optimum NAE exhibiting the lowest charge transfer resistance and the highest redox cycling performance among all NAEs. Finally, to observe the effect of their EC performance as biosensor, an EC immunoassay was performed using Interleukine-6 (IL-6), and high-density NAE has lowest a low limit of detection (LOD) of 0.45 pg/mL compared with other NAEs (medium-density: 3.91 pg/mL, low-density: 5.87 pg/mL).

Nanochannels-based self-correction electrochemical detection of mucin 1 with in-situ generated phosphomolybdic acid

Accurate detection of cancer biomarkers like mucin 1 (MUC-1) is significantly important for diagnosis and prognosis of tumor. Herein, a self-correction electrochemical immunoassay for MUC-1 detection was developed based on bioinspired nanochannels and electrochemistry of molybdophosphate. MUC-1 can be captured in the bioinspired polymer nanochannels via the formation of immunocomplex. Then, diffusion of MoO42− in the nanochannels would be sterically hindered and the in-situ generation of phosphomolybdic acid onto the electrode surface which was modified with phosphate chitosan hydrogel, would be influenced. Therefore, the two well-defined redox peaks of phosphomolybdic acid can be applied to the detection of the antigen. To improve detection sensitivity, the phosphate chitosan was incorporated with carbon nanotubes. Moreover, the occurrence in nanochannels was amplified by hybridization chain reaction, which enhanced the hindrance for transport of MoO42− by increasing steric effect, electrostatic repulsion as well as anchoring sites. Under optimized conditions, currents of the both anodic peaks on square wave voltametric voltammograms decreased linearly with the logarithm of the concentration of MUC-1 ranging from 10 fg∙mL−1 to 100 ng∙mL−1 with a detection limit of 6.0 fg∙mL−1. And thus, the detection results can be self-corrected by the results acquired from the two peaks, which had valuable implications for the reliable MUC-1 detection in disease diagnosis and therapy application.

Toward Sensitive and Reliable Immunoassays of Marine Biotoxins: From Rational Design to Food Analysis.

Marine biotoxins are metabolites produced by algae that can accumulate in shellfish or fish and enter organisms through the food chain, posing a serious threat to biological health. Therefore, accurate and rapid detection is an urgent requirement for food safety. Although various detection methods, including the mouse bioassay, liquid chromatography-mass spectrometry, and cell detection methods, and protein phosphatase inhibition assays have been developed in the past decades, the current detection methods cannot fully meet these demands. Among these methods, the outstanding immunoassay virtues of high sensitivity, reliability, and low cost are highly advantageous for marine biotoxin detection in complex samples. In this work, we review the recent 5-year progress in marine biotoxin immunodetection technologies such as optical immunoassays, electrochemical immunoassays, and piezoelectric immunoassays. With the assistance of immunoassays, the detection of food-related marine biotoxins can be implemented for ensuring public health and preventing food poisoning. In addition, the immunodetection technique platforms including lateral flow chips and microfluidic chips are also discussed. We carefully investigate the advantages and disadvantages for each immunoassay, which are compared to demonstrate the guidance for selecting appropriate immunoassays and platforms for the detection of marine biotoxins. It is expected that this review will provide insights for the further development of immunoassays and promote the rapid progress and successful translation of advanced immunoassays with food safety detection.

Electrochemical Lateral Flow Immunoassay with Built-In Electrodes for Ultrasensitive and Wireless Detection of Inflammatory Biomarkers.

Paper-based lateral flow immunoassays (LFIAs) are cost-effective, portable, and simple methods for detection of diverse analytes, which however only provide qualitative or semiquantitative results and lack sufficient sensitivity. A combination of LFIA and electrochemical detection, namely, electrochemical lateral flow immunoassay (eLFIA), enables quantitative detection of analytes with high sensitivity, but the integration of external electrodes makes the system relatively expensive and unstable. Herein, the working, counter, and reference electrodes were prepared directly on the nitrocellulose membrane using screen printing, which remarkably simplified the structure of eLFIA and decreased the cost. Moreover, a horseradish peroxidase (HRP)-based electrochemical signal amplification strategy was used for further increasing the analytical sensitivity. HRP captured on the working electrode can catalyze the oxidation of tetramethylbenzidine (TMB) to form the TMB-TMBox precipitate on the electrode surface, which as an electrochemically active product can output an amplified current for quantification. We demonstrated that the eLFIA could detect low-abundant inflammatory biomarkers in human plasma samples with limits of detection of 0.17 and 0.54 pg mL-1 for interleukin-6 and C-reactive protein, respectively. Finally, a fully portable system was fabricated by integrating eLFIA with a flexible and wireless electrochemical workstation, realizing the point-of-care detection of interleukin-6.

An electrochemical immunosensor based on MXene-GQD/AuNPs for the detection of trace amounts of CA-125 as specific tracer of ovarian cancer.

An electrochemical immunoassay system was developed to detect CA-125 using a glassy carbon electrode (GCE) modified with MXene, graphene quantum dots (GQDs), and gold nanoparticles (AuNPs). The combined MXene-GQD/AuNPs modification displayed advantageous electrochemical properties due to the synergistic effects of MXene, GQDs, and AuNPs. The MXene-GQD composite in the modified layer provided strong mechanical properties and a large specific surface area. Furthermore, the presence of AuNPs significantly improved conductivity and facilitated the binding of anti-CA-125 on the modified GCE, thereby enhancing sensitivity. Various analytical techniques such as FE-SEM and EDS were utilized to investigate the structural and morphological characteristics as well as the elemental composition. The performance of the developed immunosensor was assessed using electrochemical impedance spectroscopy (EIS), cyclic voltammetry (CV), square wave voltammetry (SWV), and differential pulse voltammetry (DPV). Under optimized conditions in a working potential range of -0.2 to 0.6V (vs. Ag/AgCl), the sensitivity, linear range (LR), limit of detection (LOD), and correlation coefficient (R2) were determined to be 315.250 µA pU.mL-1/cm2, 0.1 to 1 nU/mL, 0.075 nU/mL, and 0.9855, respectively. Thedetection of CA-125 in real samples was investigated using the developed immunoassay platform, demonstrating satisfactory results including excellent selectivity and reproducibility.

Label-free electrochemical biosensor based on dual amplification of gold nanoparticles and polycaprolactones for CEA detection

Carcinoembryonic Antigen (CEA), an acidic glycoprotein with human embryonic antigen properties, is found on the surface of cancer cells that have differentiated from endodermal cells. This paper presents a label-free electrochemical immunoassay for the dual amplification detection of CEA using gold nanoparticles loaded with polypyrrole polydopamine (Au/PPy-PDA) and polymerized polycaprolactone (Ng-PCL) prepared by ring-opening polymerization (ROP). First, the composite Au/PPy-PDA was adhered to the electrode surface. Then, gold nanoparticles form a Au–S bond with the sulfhydryl group in Apt1 to secure it on the electrode surface. Subsequently, the non-specific binding sites on the electrodes surface are closed by bovine serum albumin (BSA). Next, CEA is dropped onto the electrode surface, which is immobilized by antigen-antibody specific recognition, and the carboxyl-functionalized Apt2 forms a "sandwich structure" of antibody-antigen-antibody by specific recognition. Polymeric Ng-PCL is adhered to the electrode surface, leading to an increase in the electrochemical impedance signal, resulting in a complete chain of signal analysis. Finally, the response signal is detected by electrochemical impedance spectroscopy (EIS). Under optimal experimental conditions, the method has the advantages of high sensitivity and wide linear range (1 pg mL−1∼100 ng mL−1), and the lower limit of detection (LOD) is 0.234 pg mL−1. And it has the same high sensitivity, selectivity and interference resistance for the real samples detection. Thus, it provides a new way of thinking about biomedical and clinical diagnosis.

A plug-and-play, easy-to-manufacture fluidic accessory to significantly enhance the sensitivity of electrochemical immunoassays

Earlier access to patients’ biomarker status could transform disease management. However, gold-standard techniques such as enzyme-linked immunosorbent assays (ELISAs) are typically not deployed at the point-of-care due to their cumbersome instrumentation and complexity. Electrochemical immunosensors can be disruptive in this sector with their small size and lower cost but, without further modifications, the performance of these sensors in complex media (e.g., blood) has been limited. This paper presents a low-cost fluidic accessory fabricated using widely accessible materials and processes for boosting sensor sensitivity through confinement of the detection media next to the electrode surface. Liquid confinement first highlighted a spontaneous reaction between the pseudoreference electrode and ELISA detection substrate 3,3’,5,5’-tetramethylbenzidine (TMB) that decreases the amount of oxTMB available for detection. Different strategies are investigated to limit this and maximize reliability. Next, flow cell integration during the signal amplification step of sensor preparation was shown to substantially enhance the detection of cytokine interleukin-6 (IL-6) with the best sensitivity boost recorded for fresh human plasma (x7 increase compared to x5.8 in purified serum and x5.5 in PBS). The flow cell requires no specialized equipment and can be seamlessly integrated with commercial sensors, making an ideal companion for electrochemical signal enhancement.

Electroanalytical Platform for Rapid E. coli O157:H7 Detection in Water Samples.

There is a pressing need to enhance early detection methods of E. coli O157:H7 to mitigate the occurrence and consequences of pathogenic contamination and associated outbreaks. This study highlights the efficacy of a portable electrochemical sensing platform that operates without faradaic processes towards detecting and quantifying E. coli O157:H7. It is specifically tailored for quick identification in potable water. The assay processing time is approximately 5 min, addressing the need for swift and efficient pathogen detection. The sensing platform was constructed utilizing specific, monoclonal E. coli antibodies, based on single-capture, non-faradaic, electrochemical immunoassay principles. The E. coli sensor assay underwent testing over a wide concentration range, spanning from 10 to 105 CFU/mL, and a limit of detection (LoD) of 1 CFU/mL was demonstrated. Significantly, the sensor's performance remained consistent across studies, with both inter- and intra-study coefficients of variation consistently below 20%. To evaluate real-world feasibility, a comparative examination was performed between laboratory-based benchtop data and data obtained from the portable device. The proposed sensing platform exhibited remarkable sensitivity and selectivity, enabling the detection of minimal E. coli concentrations in potable water. This successful advancement positions it as a promising solution for prompt on-site detection, characterized by its portability and user-friendly operation. This study presents electrochemical-based sensors as significant contributors to ensuring food safety and public health. They play a crucial role in preventing the occurrence of epidemics and enhancing the supervision of water quality.

Atomically Co-dispersed nitrogen-doped carbon for sensitive electrochemical immunoassay of breast cancer biomarker CA15-3.

The development of an ultrasensitive and precise measurement of a breast cancer biomarker (cancer antigen 15-3; CA15-3) in complex human serum is essential for the early diagnosis of cancer in groups of healthy populations and the treatment of patients. However, currently available testing technologies suffer from insufficient sensitivity toward CA15-3, which severely limits early large-scale screening of breast cancer patients. We report a versatile electrochemical immunoassay method based on atomically cobalt-dispersed nitrogen-doped carbon (Co-NC)-modified disposable screen-printed carbon electrode (SPCE) with alkaline phosphatase (ALP) and its metabolite, ascorbic acid 2-phosphate (AAP), as the electrochemical labeling and redox signaling unit for sensitive detection of low-abundance CA15-3. During electrochemical detection by differential pulse voltammetry (DPV), it was found that the Co-NC-SPCE electrode did not have a current signal response to the AAP substrate; however, it had an extremely favorable response current to ascorbic acid (AA). Based on the above principle, the target CA15-3-triggered immunoassay enriched ALP-catalyzed AAP produces a large amount of AA, resulting in a significant change in the system current signal, thereby realizing the highly sensitive detection of CA15-3. Under the optimal AAP substrate concentration and ALP catalysis time, the Co-NC-SPCE-based electrochemical immunoassay demonstrated a good DPV current for CA15-3 in the assay interval of 1.0 mU/mL to 10,000 mU/mL, with a calculated limit of detection of 0.38 mU/mL. Since Co-NC-SPCE has an excellent DPV current response to AA and employs split-type scheme, the constructed electrochemical immunoassay has the merits of high preciseness and anti-interference, and its clinical diagnostic results are comparable to those of commercial kits.

Automated enzyme-linked immunosorbent assay for point-of-care COVID-19 testing

The COVID-19 pandemic pushed the demand for readily available monitoring tools, leading to the development of electrochemical point-of-care devices. This work presents an electrochemical capillary-driven immunoassay (eCaDI) device based on polyethylene terephthalate (PET) and adhesive films featuring a screen-printed carbon electrode (SPCE) for the detection of SARS-CoV-2 nucleocapsid (N) protein in clinical samples. The SPCE was modified with protein A and capture antibodies (Ab1) to ensure biomolecule immobilization and enhance sensitivity to detect N protein. The eCaDI automates an enzyme-linked immunosorbent assay (ELISA) through sequential reagent delivery, allowing for one-step sample addition. This process involves capturing the N protein by Ab1, followed by the automated delivery of HRP-labelled antibody and 3,3’,5,5’-tetramethylbenzidine (TMB) followed by electrochemical detection through TMB reduction. The SPCEs are sensitive and selective for SARS-CoV-2 N protein and stable for four weeks. Inactivated virus was used to determine a limit of detection (LOD) of 627 PFU/mL. The results obtained through immunoassay on eCADI were consistent with those of the RT-PCR method, showing a coefficient of determination (R2) of 0.816 for 27 nasal swab samples from SARS-CoV-2 infected individuals. The developed eCADI is an alternative to conventional ELISA, given the device's low cost (less than $1), low consumption of samples and reagents, low waste generation, and short analysis time (5 min). Moreover, the immunoassay has shown potential for diagnosing COVID-19 in decentralized areas.

Modelling Prospects of Bio-Electrochemical Immunosensing Platforms

Electrochemistry is a hotspot in today’s research arena. Many different domains have been extended for their role towards the Internet of Things, digital health, personalized nutrition, and/or wellness using electrochemistry. These advances have led to a substantial increase in the power and popularity of electroanalysis and its expansion into new phases and environments. The recent COVID-19 pandemic, which turned our lives upside down, has helped us to understand the need for miniaturized electrochemical diagnostic platforms. It also accelerated the role of mobile and wearable, implantable sensors as telehealth systems. The major principle behind these platforms is the role of electrochemical immunoassays, which help in overshadowing the classical gold standard methods (reverse transcriptase polymerase chain reaction) in terms of accuracy, time, manpower, and, most importantly, economics. Many research groups have endeavoured to use electrochemical and bio-electrochemical tools to overcome the limitations of classical assays (in terms of accuracy, accessibility, portability, and response time). This review mainly focuses on the electrochemical technologies used for immunosensing platforms, their fabrication requirements, mechanistic objectives, electrochemical techniques involved, and their subsequent output signal amplifications using a tagged and non-tagged system. The combination of various techniques (optical spectroscopy, Raman scattering, column chromatography, HPLC, and X-ray diffraction) has enabled the construction of high-performance electrodes. Later in the review, these combinations and their utilization will be explained in terms of their mechanistic platform along with chemical bonding and their role in signal output in the later part of article. Furthermore, the market study in terms of real prototypes will be elaborately discussed.

Rapid diagnosis of bloodstream infections using a culture-free phenotypic platform

BackgroundBloodstream infections (BSIs) are a life-threatening acute medical condition and current diagnostics for BSIs suffer from long turnaround time (TAT). Here we show the validation of a rapid detection-analysis platform (RDAP) for the diagnosis of BSIs performed on clinical blood samplesMethodsThe validation was performed on a cohort of 59 clinical blood samples, including positive culture samples, which indicated confirmed bloodstream infections, and negative culture samples. The bacteria in the positive culture samples included Gram-positive and Gram-negative pathogenic species. RDAP is based on an electrochemical sandwich immunoassay with voltage-controlled signal amplification, which provides an ultra-low limit of detection (4 CFU/mL), allowing the platform to detect and identify bacteria without requiring culture and perform phenotypic antibiotic susceptibility testing (AST) with only 1–2 h of antibiotic exposure. The preliminary diagnostic performance of RDAP was compared with that of standard commercial diagnostic technologies.ResultsUsing a typical clinical microbiology laboratory diagnostic workflow that involved sample culture, agar plating, bacteria identification using matrix-assisted laser desorption ionization time-of-flight (MALDI TOF) mass spectrometry, and AST using MicroScan as a clinical diagnostic reference, RDAP showed diagnostic accuracy of 93.3% and 95.4% for detection-identification and AST, respectively. However, RDAP provided results at least 15 h faster.ConclusionsThis study shows the preliminary feasibility of using RDAP to rapidly diagnose BSIs, including AST. Limitations and potential mitigation strategies for clinical translation of the present RDAP prototype are discussed. The results of this clinical feasibility study indicate an approach to provide near real-time diagnostic information for clinicians to significantly enhance the treatment outcome of BSIs.

Novel electrochemical platform based on C3N4-graphene composite for the detection of neuron-specific enolase as a biomarker for lung cancer

Lung cancer remains the leading cause of cancer mortality worldwide. Small cell lung cancer (SCLC) accounts for 10–15% of cases and has an overall 5-years survival rate of only 15%. Neuron-specific enolase (NSE) has been identified as a useful biomarker for early SCLC diagnosis and therapeutic monitoring. This work reports an electrochemical immunosensing platform based on a graphene-graphitic carbon nitride (g-C3N4) nanocomposite for ultrasensitive NSE detection. The g-C3N4 nanosheets and graphene nanosheets were synthesized via liquid exfoliation and integrated through self-assembly to form the nanocomposite. This nanocomposite was used to modify screen-printed carbon electrodes followed by covalent immobilization of anti-NSE antibodies. The unique properties of the graphene-g-C3N4 composite facilitated efficient antibody loading while also enhancing electron transfer efficiency and electrochemical response. Systematic optimization of experimental parameters was performed. The immunosensor exhibited a wide linear detection range of 10 pg/mL to 100 ng/mL and low limit of detection of 3 pg/mL for NSE along with excellent selectivity against interferences. Real serum matrix analysis validated the applicability of the developed platform for sensitive and accurate NSE quantifica-tion at clinically relevant levels. This novel graphene-g-C3N4 nanocomposite based electro-chemical immunoassay demonstrates great promise for early diagnosis of SCLC.