Electrochemical Biosensor Research Articles

Reduction of Losses and Wastage in Seafoods: The Role of Smart Tools and Biosensors Based on Artificial Intelligence

This paper reviews current knowledge on the role of smart tools and biosensors based on artificial intelligence in reducing seafood loss and wastage. This study shows that a variety of biosensors, categorised according to how they function, can be used to measure the quality of seafood. These include optical biosensors, enzyme-based biosensors, immunosensors, microbial biosensors, DNA-based biosensors, electrochemical biosensors, optical biosensors, tissue-based biosensors, and piezoelectric biosensors. Among these biosensors, optical biosensors, electrochemical biosensors, and mechanical biosensors are the most significant. Again, this study report that, for seafood traceability and management, a variety of smart solutions including blockchain technology, quick response (QR) codes, data analytics, digital twins, and radio frequency identification (RFID) tags can be utilised. Catch data, vessel tracking data, and data from the processing plant are some of the different data sources that can be utilised to trace seafood products. Artificial intelligence tools like neural networks, deep learning, machine learning, and others can be used to forecast and improve seafood quality. It is crucial to study the development of biosensors that can properly identify the earliest signs of seafood contamination or rotting.

Novel MoS2-decorated Cu2O hybrid nanoparticles for enhanced non-enzymatic electrochemical cholesterol detection

Precise identification of cholesterol levels is crucial for the early diagnosis of cardiovascular risk factors. This paper presents a novel approach for cholesterol detection that circumvents the reliance on enzymatic processes. Leveraging the unique properties of advanced materials and electrochemical principles, our non-enzymatic approach demonstrates enhanced sensitivity, specificity, and limit of detection in cholesterol analysis. A non-enzymatic electrochemical biosensor for Cholesterol, employing a nanohybrid comprising Cu2O nanoparticles decorated with MoS2, is presented. The cyclic voltammetry (CV), differential pulse voltammetry (DPV), and amperometry techniques were employed to investigate the electrochemical behaviour of the glassy carbon electrode modified with the Cu2O/MoS2 nanohybrid. The modified electrode exhibited an excellent sensitivity of 111.74 μA μM−1 cm−2 through the CV method and showcased a low detection limit of 2.18 μM and an expansive linear range spanning 0.1–180 μM when employing the DPV method. The electrode also showed good selectivity to various interfering components in 0.1 M NaOH and a satisfied stability of about 15 days at room temperature. The study demonstrates the potential for broader applications in clinical diagnostics and monitoring cardiovascular health, paving the way for a paradigm shift in cholesterol detection methodologies and offering a more efficient and cost-effective alternative to traditional enzymatic assays.

A novel electrochemical biosensor based on SRCA-NEMA-G-quadruplex for sensitive detection of Listeria monocytogenes in food

Listeria monocytogenes (L. monocytogenes), as one of the most common foodborne pathogens, has caused many foodborne diseases, posing a major threat to food and public health safety. Therefore, a novel electrochemical biosensor was proposed based on the combination of saltatory rolling circle amplification (SRCA) and nicking enzyme mediated amplification (NEMA) for detecting Listeria monocytogenes. When L. monocytogenes was present, abundant double-stranded DNA (dsDNA) were produced by SRCA reaction. SRCA products could be specifically recognized and cleaved by Nb.BsmI. SRCA-NEMA was performed to generate abundant single-stranded DNA (ssDNA) with the action of Bst DNA polymerase and Nb.BsmI. The hydrosulfuryl-modified hairpin DNA (HS-hpDNA) was immobilized on the electrode surface and G-quadruplex was used as signaling molecule. The current signal increased with target DNA concentration and the ratio of the current could be calculated as the result. Subsequently, the detection linear range for L. monocytogenes genomic DNA was 5.4 × 100-5.4 × 107 fg/μL, with the detection limit of 2.13 fg/μL. In comparison with ISO 11290-1:2017 method, the relative sensitivity, relative specificity and relative accuracy of the developed method were 100 %, 97.8 % and 98 %, respectively. Overall, the developed method provides an outstanding platform for sensitive detection of L. monocytogenes in food.

Electrochemical biosensors for pathogenic microorganisms detection based on recognition elements.

The worldwide spread of pathogenic microorganisms poses a significant risk to human health. Electrochemical biosensors have emerged as dependable analytical tools for the point-of-care detection of pathogens and can effectively compensate for the limitations of conventional techniques. Real-time analysis, high throughput, portability, and rapidity make them pioneering tools for on-site detection of pathogens. Herein, this work comprehensively reviews the recent advances in electrochemical biosensors for pathogen detection, focusing on those based on the classification of recognition elements, and summarizes their principles, current challenges, and prospects. This review was conducted by a systematic search of PubMed and Web of Science databases to obtain relevant literature and construct a basic framework. A total of 171 publications were included after online screening and data extraction to obtain information of the research advances in electrochemical biosensors for pathogen detection. According to the findings, the research of electrochemical biosensors in pathogen detection has been increasing yearly in the past 3years, which has a broad development prospect, but most of the biosensors have performance or economic limitations and are still in the primary stage. Therefore, significant research and funding are required to fuel the rapid development of electrochemical biosensors. The overview comprehensively evaluates the recent advances in different types of electrochemical biosensors utilized in pathogen detection, with a view to providing insights into future research directions in biosensors.

Electrochemical biosensor for rapid and sensitive monitoring of sulfadimethoxine based on nanoporous carbon and aptamer system

In this study, a straightforward electrochemical aptasensor was developed to detect sulfadimethoxine (SDM). It included a glassy carbon electrode decorated by boron nitride quantum dots (BNQDs) and aptamer-functionalized nanoporous carbon (APT/CZ). CZ was first synthesized by calcinating a zeolitic imidazolate framework (ZIF-8). Then, the electroactive dye methylene blue (MB) was entrapped inside its pores. By attaching aptamer to the CZ surface, APT/CZ acted as a bioguard, which prevented the MB release. Therefore, the electrochemical signal of the entrapped MB was high in the absence of SDM. Introducing SDM caused the conformation of aptamers to change, and a large number of MB was released, which was removed by washing. Therefore, the detection strategy was done based on the change in the electrochemical signal intensity of MB. The aptasensor was applied to detect SDM at a concentration range of 10−17 to 10−7 M with a detection limit of 3.6 × 10−18 M.

Stretchable Nanofiber-Based Felt as a String Electrode for Potential Use in Wearable Glucose Biosensors.

Nanofiber technology is leading the revolution of wearable technology and provides a unique capability to fabricate smart textiles. With the novel fabrication technique of electrospinning, nanofibers can be fabricated and then manufactured into a durable conductive string for the application of smart textiles. This paper presents an electrospun nanofiber mesh-based (NF-Felt) string electrode with a conducting polymer coating for an electrochemical enzymatic glucose sensor. The surface area of a nanofiber matrix is a key physical property for enhanced glucose oxidase (GOx) enzyme binding for the development of an electrochemical biosensor. A morphological characterization of the NF-Felt string electrode was performed using scanning electron microscopy (SEM) and compared with a commercially available cotton-polyester (Cot-Pol) string coated with the same conducting polymer. The results from stress-strain testing demonstrated high stretchability of the NF-Felt string. Also, the electrochemical characterization results showed that the NF-Felt string electrode was able to detect a glucose concentration in the range between 0.0 mM and 30.0 mM with a sensitivity of 37.4 μA/mM·g and a detection limit of 3.31 mM. Overall, with better electrochemical performance and incredible flexibility, the NF-Felt-based string electrode is potentially more suitable for designing wearable biosensors for the detection of glucose in sweat.

A trivalent aptasensor by using DNA tetrahedron as scaffold for label-free determination of antibiotics

Owing to advantage in high sensitivity and fast response, aptamer based electrochemical biosensors have attracted much more attention. However, inappropriate interfacial engineering strategy leads to poor recognition performance, which ascribe to the following factors of immobilized oligonucleotide strand including steric hindrance, interchain entanglement, and unfavorable conformation. In this work, we proposed a DNA tetrahedron based diblock aptamer immobilized strategy for the construction of label-free electrochemical biosensor. The diblock aptamer sequence is composite of T-rich anchor domain and recognition domain, where T-rich domain enabling anchored on the edge of DNA tetrahedron via Hoogsteen hydrogen bond at neutral condition. The DNA tetrahedron scaffold offers an appropriate lateral space for target recognition of diblock aptamer. More importantly, this trivalent aptamer recognition interface can be regenerated by simply adjusting the pH environment to alkaline, resulting in the dissociation of diblock aptamer. Under the optimum condition, proposed electrochemical aptasensor manifested a satisfied sensitivity for aminoglycosides antibiotic, kanamycin with a limit of detection of 0.69 nM, which is 45-fold lower than traditional Au–S immobilization strategy. Moreover, the proposed aptasensor had also successfully been extended to ampicillin detection by changing the sequence of recognition domain in diblock aptamer. This work paves a new way for the rational design of aptamer-based electrochemical sensor.

Nucleic Acid-based Electrochemical Sensors Facilitate the Study of DNA Binding by Platinum (II)-based Antineoplastics.

DNA crosslinking agents such as cisplatin and related platinum(II) analogs are effective drugs to treat solid tumors. However, these therapeutics can cause high toxicity in the body, and tumors can develop resistance to them. To develop less toxic and more effective DNA crosslinkers, medicinal chemists have focused on tuning the ligands in square planar platinum(II) complexes to modulate their bioavailability, targeted cell penetration, and DNA binding rates. Unfortunately, linking in vitro DNA binding capacity of DNA crosslinkers with their in vivo efficacy has proven challenging. Here we report an electrochemical biosensor strategy that allows the study of platinum(II)-DNA binding in real time. Our biosensors contain a purine-rich deoxynucleotide sequence, T6 (AG)10 , modified with a 5' hexylthiol linker for easy self-assembly onto gold electrodes. The 3' terminus is functionalized with the redox reporter methylene blue. Electron transfer from methylene blue to the sensor is a function of platinum(II) compound concentration and reaction time. Using these biosensors, we resolve DNA binding mechanisms including monovalent and bivalent binding, as well as base stacking. Our approach can measure DNA binding kinetics in buffers and in 50 % serum, offering a single-step, real-time approach to screen therapeutic compounds during drug development.

Electrochemical biosensor for the evaluation of monoclonal antibodies targeting the N protein of SARS-CoV-2 virus

The emergence of COVID-19 caused by the coronavirus SARS-CoV-2 has prompted a global pandemic that requires continuous research and monitoring. This study presents a design of an electrochemical biosensing platform suitable for the evaluation of monoclonal antibodies targeting the SARS-CoV-2 nucleocapsid (N) protein. Screen-printed carbon electrodes (SPCE) modified with gold nanostructures (AuNS) were applied to design a versatile and sensitive sensing platform. Electrochemical techniques, including electrochemical impedance spectroscopy (EIS) and square wave voltammetry (SWV), were used to investigate the interactions between immobilised recombinant N (rN) protein and several monoclonal antibodies (mAbs). The electrochemical characterisation of SPCE/AuNS/rN demonstrated a successful immobilisation of rN, enhancing the electron transfer kinetics. Affinity interactions between immobilised rN and four mAbs (mAb-4B3, mAb-4G6, mAb-12B2, and mAb-1G5) were explored. Although mAb-4B3 showed some non-linearity, the other monoclonal antibodies exhibited specific and well-defined interactions followed by the formation of an immune complex. The biosensing platform demonstrated high sensitivity in the linear range (LR) from 0.2 nM to 1 nM with limits of detection (LOD) ranging from 0.012 nM to 0.016 nM for mAb-4G6, mAb-12B2, and mAb-1G5 and limits of quantification (LOQ) values ranging from 0.035 nM to 0.139 nM, as determined by both EIS and SWV methods. These results highlight the system's potential for precise and selective detection of monoclonal antibodies specific to the rN. This electrochemical biosensing platform provides a promising route for the sensitive and accurate detection of monoclonal antibodies specific to the rN protein.

Multiplex electrochemical sensing platforms for the detection of breast cancer biomarkers.

Herein, advancements in electroanalytical devices for the simultaneous detection of diverse breast cancer (BC) markers are demonstrated. This article identifies several important areas of exploration for electrochemical diagnostics and highlights important factors that are pivotal for the successful deployment of novel bioanalytical devices. We have highlighted that the limits of detection (LOD) reported for the multiplex electrochemical biosensor can surpass the sensitivity displayed by current clinical standards such as ELISA, FISH, and PCR. HER-2; a breast cancer marker characterised by increased metastatic potential, more aggressive development, and poor clinical outcomes; can be sensed with a LOD of 0.5 ng/ml using electrochemical multiplex platforms, which falls within the range of that measured by ELISA (from picogram/ml to nanogram/ml). Electrochemical multiplex biosensors are reported with detection limits of 0.53 ng/ml and 0.21 U/ml for MUC-1 and CA 15-3, respectively, or 5.8 × 10-3 U/ml for CA 15-3 alone. The sensitivity of electrochemical assays is improved when compared to conventional analysis of MUC-1 protein which is detected at 11-12 ng/ml, and ≤30 U/ml for CA 15-3 in the current clinical blood tests. The LOD for micro-ribonucleic acid (miRNA) biomarkers analyzed by electrochemical multiplex assays were all notedly superior at 9.79 × 10-16 M, 3.58 × 10-15 M, and 2.54 × 10-16 M for miRNA-155, miRNA-21, and miRNA-16, respectively. The dogma in miRNA testing is the qRT-PCR method, which reports ranges in the ng/ml level for the same miRNAs. Breast cancer exosomes, which are being explored as a new frontier of biosensing, have been detected electrochemically with an LOD of 103-108 particles/mL and can exceed detection limits seen by the tracking and analysis of nanoparticles (∼ 107 particles/ml), flow cytometry, Western blotting and ELISA, etc. A range of concentration at 78-5,000 pg/ml for RANKL and 16-1,000 pg/ml for TNF is reported for ELISA assay while LOD values of 2.6 and 3.0 pg/ml for RANKL and TNF, respectively, are demonstrated by the electrochemical dual immunoassay platform. Finally, EGFR and VEGF markers can be quantified at much lower concentrations (0.01 and 0.005 pg/ml for EGFR and VEGF, respectively) as compared to their ELISA assays (EGRF at 0.31-20 ng/ml and VEGF at 31.3-2,000 pg/ml). In this study we hope to answer several questions: (1) Are the limits of detection (LODs) reported for multiplex electrochemical biosensors of clinical relevance and how do they compare to well-established methods like ELISA, FISH, or PCR? (2) Can a single sensor electrode be used for the detection of multiple markers from one blood drop? (3) What mechanism of electrochemical biosensing is the most promising, and what technological advancements are needed to utilize these devices for multiplex POC detection? (4) Can nanotechnology advance the sensitive and selective diagnostics of multiple BC biomarkers? (5) Are there preferred receptors (antibody, nucleic acid or their combinations) and preferred biosensor designs (complementary methods, sandwich-type protocols, antibody/aptamer concept, label-free protocol)? (6) Why are we still without FDA-approved electrochemical multiplex devices for BC screening?

Multiplex signal amplification for ultrasensitive CRP assay via integrated electrochemical biosensor array using MOF-derived carbon material and aptamers

Accurate and precise detection of disease-associated proteins, such as C-reactive protein (CRP), remains a challenge in biosensor development. Herein, we present a novel approach－an integrated disposable aptasensor array－designed for precise, ultra-sensitive, and parallel detection of CRP in plasma samples. This integrated biosensing array platform enables multiplex parallel testing, ensuring the accuracy and reliability in sample analysis. The ultra-sensitivity of this biosensor is achieved through multiplex signal amplification. Leveraging the superior conductivity and extensive surface area of MOF-derived nanoporous carbon material (CMOF), the biosensor enhances recognition elements (aptamers) by catalyzing the horseradish peroxidase (HRP) label enzyme reaction to multiply the number of probe molecules. Optimized conditions yielded exceptional performance, exhibiting high accuracy (relative standard deviation, RSD≤10.0 %), a low detection limit (0.3 pg/mL, S/N = 3), ultra-sensitivity (0.16 μA/ng mL−1 mm−2), and a rapid response (seven parallel tests within 60 min). Importantly, this multi-unit integrated disposable aptasensor array accurately quantified CRP in human serum, demonstrating comparable results to commercial enzyme–linked immunosorbent assay (ELISA). This technology showcases promise for detecting various biomarkers using a unified approach, presenting an appealing strategy for early disease diagnosis and biological analysis.

Electronic Tongue Based on ZnO/ITO@glass for Electrochemical Monitoring of Spiciness Levels.

Capsaicin, a chemical compound present in chili peppers, is widely acknowledged as the main contributor to the spicy and hot sensations encountered during consumption. Elevated levels of capsaicin can result in meals being excessively spicy, potentially leading to health issues, such as skin burning, irritation, increased heart rate and circulation, and discomfort in the gastrointestinal system and even inducing nausea or diarrhea. The level of spiciness that individuals can tolerate may vary, so what may be considered incredibly hot for one person could be mild for another. To ensure food safety, human healthcare, regulatory compliance, and quality control in spicy food products, capsaicin levels must be measured. For these purposes, a reliable and stable sensor is required to quantify the capsaicin level. To leverage the effect of zinc oxide (ZnO), herein, we demonstrated the one-step fabrication process of an electronic tongue (E-Tongue) based on an electrochemical biosensor for the determination of capsaicin. ZnO was electrodeposited on the indium tin oxide (ITO) surface. The biosensor demonstrated the two notable linear ranges from 0.01 to 50 μM and from 50 to 500 μM with a limit of detection (LOD) of 2.1 nM. The present study also included the analysis of real samples, such as green chilis, red chili powder, and dried red chilis, to evaluate their spiciness levels. Furthermore, the E-Tongue exhibited notable degrees of sensitivity, selectivity, and long-term stability for a duration of more than a month. The development of an E-Tongue for capsaicin real-time monitoring as a point-of-care (POC) device has the potential to impact various industries and improve safety, product quality, and healthcare outcomes.

Electrochemical flow-through biosensors based on microfiber enzymatic filter discs placed at printed electrodes

A new type of electrochemical biosensors in a flow injection system with printed electrodes were developed and tested. A filter disc (7 mm diameter) with immobilized enzyme was placed at the printed electrode. This conception combines the advantages of biosensors with a bioreceptor at the electrode surface and systems with spatially separated enzymatic and detection parts. Filters of different composition (glass, quartz, and cellulose), thickness, porosity, and ways of binding enzyme to their surface were tested. Only covalent bonds throughout a filter-aminosilane-glutaraldehyde-enzyme chain ensured a long-time and reproducible biosensor response. The developed method of biosensor preparation has been successfully applied to enzymes glucose oxidase, laccase and choline oxidase. The dependences of peak current on detection potential, flow rate, injection volume, analyte concentration as well as biosensor lifetime and reproducibility were investigated for glucose oxidase biosensor. The sensitivity of measurements was two or more times higher than that of biosensor with a mini-reactor filled by powder with immobilized enzyme. The developed biosensor with laccase was tested by determining dopamine in the pharmaceutical infusion product Tensamin®. Results of the analysis (40.0 ± 0.7 mg mL−1, SD = 0.8 mg mL−1, RSD = 1.85 %, N = 11) show a good agreement with the manufacturer's declared value.

Electrochemical Acetylcholinesterase Sensors for Anti-Alzheimer's Disease Drug Determination.

Neurodegenerative diseases and Alzheimer's disease (AD), as one of the most common causes of dementia, result in progressive losses of cholinergic neurons and a reduction in the presynaptic markers of the cholinergic system. These consequences can be compensated by the inhibition of acetylcholinesterase (AChE) followed by a decrease in the rate of acetylcholine hydrolysis. For this reason, anticholinesterase drugs with reversible inhibition effects are applied for the administration of neurodegenerative diseases. Their overdosage, variation in efficiency and recommendation of an individual daily dose require simple and reliable measurement devices capable of the assessment of the drug concentration in biological fluids and medications. In this review, the performance of electrochemical biosensors utilizing immobilized cholinesterases is considered to show their advantages and drawbacks in the determination of anticholinesterase drugs. In addition, common drugs applied in treating neurodegenerative diseases are briefly characterized. The immobilization of enzymes, nature of the signal recorded and its dependence on the transducer modification are considered and the analytical characteristics of appropriate biosensors are summarized for donepezil, huperzine A, rivastigmine, eserine and galantamine as common anti-dementia drugs. Finally, the prospects for the application of AChE-based biosensors in clinical practice are discussed.

Porous membranes integrated into electrochemical systems for bioanalysis

AbstractPorous membranes have emerged as promising platforms for bioanalysis because of their unique properties including high surface area, selective permeability, and compatibility with electrochemical techniques. This minireview presents an overview of the development and applications of porous membrane‐based electrochemical systems for bioanalysis. First, we discuss the existing fabrication methods for porous membranes. Next, we summarize electrochemical detection strategies for bioanalysis using porous membranes. Electrochemical biosensors and cell chips fabricated from porous membranes are discussed as well. Furthermore, porous micro‐/nanoneedle devices for bioapplications are described. Finally, the utilization of scanning electrochemical microscopy for cell analysis on porous membranes and electrochemiluminescence sensors is demonstrated. Future perspectives of the described membrane detection strategies and devices are outlined in each section. This work can help enhance the performance of porous membrane‐based electrochemical systems and expand the range of their potential applications.

A direct electrochemical biosensor for rapid glucose detection based on nitrogen-doped carbon nanocages

A direct electrochemical biosensor for rapid glucose detection based on nitrogen-doped carbon nanocages

Effect of Modification of a Laccase-Based Electrochemical Biosensor with Carbon Nanotubes on Signal Separation of Dihydroxybenzene Isomers

Effect of Modification of a Laccase-Based Electrochemical Biosensor with Carbon Nanotubes on Signal Separation of Dihydroxybenzene Isomers

Disposable Polyaniline/m-Phenylenediamine-Based Electrochemical Lactate Biosensor for Early Sepsis Diagnosis.

Lactate serves as a crucial biomarker that indicates sepsis assessment in critically ill patients. A rapid, accurate, and portable analytical device for lactate detection is required. This work developed a stepwise polyurethane-polyaniline-m-phenylenediamine via a layer-by-layer based electrochemical biosensor, using a screen-printed gold electrode for lactate determination in blood samples. The developed lactate biosensor was electrochemically fabricated with layers of m-phenylenediamine, polyaniline, a crosslinking of a small amount of lactate oxidase via glutaraldehyde, and polyurethane as an outer membrane. The lactate determination using amperometry revealed the biosensor's performance with a wide linear range of 0.20-5.0 mmol L-1, a sensitivity of 12.17 ± 0.02 µA·mmol-1·L·cm-2, and a detection limit of 7.9 µmol L-1. The developed biosensor exhibited a fast response time of 5 s, high selectivity, excellent long-term storage stability over 10 weeks, and good reproducibility with 3.74% RSD. Additionally, the determination of lactate in human blood plasma using the developed lactate biosensor was examined. The results were in agreement with the enzymatic colorimetric gold standard method (p > 0.05). Our developed biosensor provides efficiency, reliability, and is a great potential tool for advancing lactate point-of-care testing applications in the early diagnosis of sepsis.

Adaptive Epidermal Bioelectronics by Highly Breathable and Stretchable Metal Nanowire Bioelectrodes on Electrospun Nanofiber Membrane

AbstractThe potential of the electrospun nanofiber membrane (ENM)‐based soft electronics in epidermal bioelectronics has gained huge attention with their conformal compatibility with the human body and associated performance improvements. This study presents a novel filtration‐based direct local nanowire patterning method on the ENM using dispenser systems, aiming to fabricate stretchable, breathable, and highly conductive epidermal electronics harnessing various types of metal nanowires, including Ag, Ag@Au core–shell, and Ag@(Au–Pt) core–shell nanowires. By utilizing capillary force from a support bed beneath the ENM, efficient fluid flow can be achieved, eliminating the requirement for expensive vacuum equipment typically employed in filtration processes. In the postprocessing phase, the photothermal effect of a laser is harnessed to improve the mechanical stability of the nanowire–ENM interface. The maskless fabrication process is instrumental in crafting epidermal bioelectronics in that the design can be spontaneously replicated according to diverse human body geometries in situ. The selective insulation process can be also executed with the same dispenser system, streamlining the overall fabrication system. Applications are demonstrated showcasing the advantages of the presented fabrication system and the resulting devices, including an in vivo epicardial signal recording electrode, an epidermal electrochemical biosensor, and a customized epidermal electromyography (EMG)‐based human–machine interface (HMI).

Electrochemical biosensor based on an atomic layered composite of graphene oxide/graphene as an electrode material towards selective and sensitive detection of miRNA-21

Development of an efficient technique towards the detection of miRNAs is highly significant for the timely diagnosis and prognosis of cancers. In this study, we utilized the chemical vapor deposition (CVD) technique to grow bilayer graphene (BLG) on copper foil and subsequently transferred the BLG to fluorine doped tin oxide (FTO) substrate via a facile wet transfer technique. The upper layer of the BLG was modified to graphene oxide (GO) with a low damage plasma treatment (LDPT) process to serve as a biomolecule receiving layer and the lower graphene layer (G) serves as an electron transport layer. The developed GO/G/FTO platform offers superior sensing ability towards the miRNA-21. The results demonstrated that the proposed GO/G/FTO substrate detect the miRNA-21 with a wide linear range (10 fM ∼ 1 nM), low limit of detection (3.18 fM), high sensitivity (0.6 mApM−1cm−2), selectivity and stability via the differential pulse voltammetry method. Additionally, the sensors can efficiently discriminate the target miRNA-21 from its complementary base mismatched counterparts. The proposed approach serves as an excellent tool for the early diagnosis of diseases.