Design of a rapid electrochemical biosensor based on MXene/Pt/C nanocomposite and DNA/RNA hybridization for the detection of COVID-19.

Since the rapid spread of the SARS-CoV-2 (2019), the need for early diagnostic techniques to control this pandemic has been highlighted. Diagnostic methods based on virus replication, such as RT-PCR, are exceedingly time-consuming and expensive. As a result, a rapid and accurate electrochemical test which is both available and cost-effective was designed in this study. MXene nanosheets (Ti3C2Tx) and carbon platinum (Pt/C) were employed to amplify the signal of this biosensor upon hybridization reaction of the DNA probe and the virus's specific oligonucleotide target in the RdRp gene region. By the differential pulse voltammetry (DPV) technique, the calibration curve was obtained for the target with varying concentrations ranging from 1 aM to 100 nM. Due to the increase in the concentration of the oligonucleotide target, the signal of DPV increased with a positive slope and a correlation coefficient of 0.9977. Therefore, at least a limit of detection (LOD) was obtained 0.4 aM. Furthermore, the specificity and sensitivity of the sensors were evaluated with 192 clinical samples with positive and negative RT-PCR tests, which revealed 100% accuracy and sensitivity, 97.87% specificity and limit of quantification (LOQ) of 60 copies/mL. Besides, various matrices such as saliva, nasopharyngeal swabs, and serum were assessed for detecting SARS-CoV-2 infection by the developed biosensor, indicating that this biosensor has the potential to be used for rapid Covid-19 test detection.

Sensitive Detection of SARS-CoV-2 Spike Protein based on Electrochemical Impedance Spectroscopy of Fe3O4@SiO2-Au/GCE Biosensor

Highly contagious COVID-19 disease is caused by a novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which poses a serious threat to global public health. Therefore, the development of a fast and reliable method for the detection of SARS-CoV-2 is an urgent research need. The Fe3O4@SiO2-Au is enriched with a variety of functional groups, which can be used to fabricate a sensitive electrochemical biosensor by biofunctionalization with angiotensin-converting enzyme 2 (ACE2). Accordingly, we developed a novel electrochemical sensor by chemically modifying a glassy carbon electrode (GCE) with Fe3O4@SiO2-Au nanocomposites (hereafter Fe3O4@SiO2-Au/GCE) for the rapid detection of S-protein spiked SARS-CoV-2 by electrochemical impedance spectroscopy (EIS). The new electrochemical sensor has a low limit detection (viz., 4.78 pg/mL) and a wide linear dynamic range (viz., 0.1 ng/mL to 10 μg/mL) for detecting the EIS response signal of S-protein. The robust Fe3O4@SiO2-Au/GCE biosensor has high selectivity, stability, and reproducibility for the detection of S-protein with good recovery of saliva samples.

Importance of IP-10 as a biomarker of host immune response: Critical perspective as a target for biosensing

While the exploration into biomolecules for diagnostic and prognostic devices continues to develop, many molecules continue to be examined for individual diseases or treatments. Consequently, it can be difficult to fully understand the scope of one individual molecule's current and potential clinical utilization. The scope of this study aimed to assess the potential of Interferon Gamma-induced Protein 10 (IP-10) as a biomarker in a wide variety of diseases, both as a main and supplemental indicator of disease infection and progression. IP-10 is a chemokine secreted in response to IFN-gamma playing a major role in the activation and regulation of inflammatory and immune responses within the body. Currently, IP-10 has displayed potential application in diseases such as COVID-19, tuberculosis, sepsis, Kawasaki disease, cancer, and many more. Molecular assays developed for the detection of IP-10 take longer testing time, sophisticated instrument utilization, and need more sample volumes. These cannot be utilized for bedside patient monitoring during the illness state of the patient. Biosensing tools are alternative methods used at clinical sites due to their rapid results. Though many types of sensing mechanisms established for the detection of disease biomarkers such as optical, piezoelectric sensors, and electrochemical biosensors are far beyond the other sensing methods due to their ease of mechanism, rapid results, and portable nature. IP-10 has been a promising biomarker in different diseases, evaluation of IP-10 levels at different time points of treatments is necessary. To achieve this, current conventional methods cannot be used and thus a portable device that provides rapid results is in demand. Such point-of-care (POC) device development for IP-10 analysis is very crucial in the current scenario. Beyond this, the clarification of its physiological role in healthy and infected individuals could allow for more proper utilization in clinical diagnoses, prognoses, treatment monitoring, and more. Overall, this study was developed to summarize the associations currently created between levels of IP-10 and other biomolecules and diseases.Copyright © 2023 The Author(s)

Synthesis and characterization of Au-decorated graphene oxide nanocomposite for magneto-electrochemical detection of SARS-CoV-2 nucleocapsid gene.

Fast and effective diagnosis is the first step in monitoring the current coronavirus 2 (CoV-2) pandemic. Herein, we establish a simple and sensitive electrochemical assay using magnetic nanocomposite and DNA sandwich probes to rapidly quantify the CoV-2 nucleocapsid (N) gene down to the 0.37 fM level. This assay uses a pair of specific DNA probes. The capture probe is covalently conjugated to Au-decorated magnetic reduced graphene oxide (AMrGO) nanocomposite for efficiently capturing target RNA. In contrast, the detection probe is linked to peroxidase for signal amplification. The probes target the COV-2 gene, allowing for specific magnetic separation, enzymatic signal amplification, and subsequent generation of voltammetric current with a total assay time of 45 min. The developed biosensor has high selectivity and can discriminate non-specific gene sequences. Synthetic COV-2 N-gene can be detected efficiently in serum and saliva, while 1-bp mismatch gene yielded a low response. The performance of the genosensor was good in an extensive linear range of 5 aM-50 pM. For synthetic N-gene, we achieved the detection limit of 0.37, 0.33, and 0.19 fM in human saliva, urine, and serum. This simple, selective, and sensitive genosensor could have various genetics-based biosensing and diagnostic applications.

Development of Aptasensor for the Diagnosis of Covid-19

Objectives: The objective is to develop a low-cost, practical, portable aptasensor platform for the diagnosis of COVID-19. Materials -Methods: Amino-terminated aptamers to be used for the design of an aptasensor were synthesized by SELEX method, and interaction of aptamers with SARS-CoV-2 S1 protein was investigated by isothermal titration calorimetry (ITC). Gold electrodes were used to design the biosensor platform. After the electrode surface was functionalized with cysteamine, the amino-terminated aptamer was conjugated to the surface via glutaraldehyde crosslinker. Then, the surface characterization and analytical parameters of the designed sensing platform were determined by adding commercial S1 proteins on the surface using differential pulse voltammetry (DPV), cyclic voltammetry (CV) and impedance spectroscopy (EIS). To evaluate the working performance of the system, S1 proteins were added to the synthetic serum samples using the standard addition method and the measurements were repeated. Result(s): Surface characterization of the platform designed with EIS and CV measurements was performed and it was found that the modification was successfully performed. In addition, DPV results and analytical parameters of the platform (calibration plot, limit of detection(LOD) , repeatability, coefficient of variation) were determined and the working performance of system was evaluated. Moreover, working performance of the biosensor in real samples and its specificity for COVID -19 were determined by experiments with synthetic serum and influenza A and B proteins. Conclusion(s): According the results, the system has potential to be used for the detection of COVID -19, and also it can be rapidly adapted in different pandemic situations that may occur in the future.

A surface molecularly imprinted electrochemical biosensor for the detection of SARS-CoV-2 spike protein by using Cu7S4-Au as built-in probe.

Sensitive detection of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) spike protein (S protein) is of significant clinical importance in the diagnosis of COVID-19 pandemic. In this work, a surface molecularly imprinted (SMI) electrochemical biosensor is fabricated for the detection of SARS-CoV-2 S protein. Cu7S4-Au is used as the built-in probe and modified on the surface of a screen-printed carbon electrode (SPCE). 4-Mercaptophenylboric acid (4-MPBA) is anchored to the surface of the Cu7S4-Au through Au-SH bonds, which can be used for the immobilization of the SARS-CoV-2 S protein template through boronate ester bonds. After that, 3-aminophenylboronic acid (3-APBA) is electropolymerized on the electrode surface and used as the molecularly imprinted polymers (MIPs). The SMI electrochemical biosensor is obtained after the elution of the SARS-CoV-2 S protein template with an acidic solution by the dissociation of the boronate ester bonds, which can be utilized for sensitive detection of the SARS-CoV-2 S protein. The developed SMI electrochemical biosensor displays high specificity, reproducibility and stability, which might be a potential and promising candidate for the clinical diagnosis of COVID-19.

Functional Nanomaterials for Electrochemical SRAS-CoV-2 Biosensors: a Review

The pandemic outbreak of COVID-19 has posed a threat to public health globally, and rapid and accurate identification of the viruses is crucial for controlling COVID-19. In recent years, nanomaterial-based electrochemical sensing techniques hold immense potential for molecular diagnosis with high sensitivity and specificity. In this review, we briefly introduced the structural characteristics and routine detection methods of SARS-CoV-2, then summarized the associated properties and mechanisms of the electrochemical biosensing methods. On the above basis, the research progress of electrochemical biosensors based on gold nanomaterials, oxide nanomaterials, carbon-based nanomaterials and other nanomaterials for rapid and accurate detection of virus were reviewed. Finally, the future applications of nanomaterial-based biosensors for biomolecular diagnostics were pointed out.

CRISPR-Cas assisted diagnostics: A broad application biosensing approach

In addition to its remarkable genome editing capability, the CRISPR-Cas system has proven to be very effective in many fields of application, including the biosensing of pathogenic infections, mutagenic defects, or early cancer diagnosis. Thanks to their many advantages in terms of simplicity, efficiency, and reduced time, several CRISPR-Cas systems have been described for the design of sensitive and selective analytical tools, paving the way for the development and further commercialization of next-generation diagnostics. However, CRISPR-Cas-based biosensors still need further research efforts to improve some drawbacks, such as the need for target amplification, low reproducibility, and lack of knowledge of exploited element robustness. This review aims to describe the latest trends in the design of CRISPR-Cas biosensing technologies to better highlight the insights of their advantages and to point out the limitations that still need to be overcome for their future market entry as medical diagnostics.Copyright © 2023 Elsevier B.V.

Functional Nanomaterials for Electrochemical SRAS-CoV-2 Biosensors: a Review

The pandemic outbreak of COVID-19 has posed a threat to public health globally, and rapid and accurate identification of the viruses is crucial for controlling COVID-19. In recent years, nanomaterial-based electrochemical sensing techniques hold immense potential for molecular diagnosis with high sensitivity and specificity. In this review, we briefly introduced the structural characteristics and routine detection methods of SARS-CoV-2, then summarized the associated properties and mechanisms of the electrochemical biosensing methods. On the above basis, the research progress of electrochemical biosensors based on gold nanomaterials, oxide nanomaterials, carbon-based nanomaterials and other nanomaterials for rapid and accurate detection of virus were reviewed. Finally, the future applications of nanomaterial-based biosensors for biomolecular diagnostics were pointed out. © 2023 Science Press. All rights reserved.

CRISPR-Cas13a-powered electrochemical biosensor for the detection of the L452R mutation in clinical samples of SARS-CoV-2 variants.

Since the end of 2019, a highly contagious disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has deprived numerous lives worldwide, called COVID-19. Up to date, omicron is the latest variant of concern, and BA.5 is replacing the BA.2 variant to become the main subtype rampaging worldwide. These subtypes harbor an L452R mutation, which increases their transmissibility among vaccinated people. Current methods for identifying SARS-CoV-2 variants are mainly based on polymerase chain reaction (PCR) followed by gene sequencing, making time-consuming processes and expensive instrumentation indispensable. In this study, we developed a rapid and ultrasensitive electrochemical biosensor to achieve the goals of high sensitivity, the ability of distinguishing the variants, and the direct detection of RNAs from viruses simultaneously. We used electrodes made of MXene-AuNP (gold nanoparticle) composites for improved sensitivity and the CRISPR/Cas13a system for high specificity in detecting the single-base L452R mutation in RNAs and clinical samples. Our biosensor will be an excellent supplement to the RT-qPCR method enabling the early diagnosis and quick distinguishment of SARS-CoV-2 Omicron BA.5 and BA.2 variants and more potential variants that might arise in the future.

Functional Nanomaterials for Electrochemical SRAS-CoV-2 Biosensors: a Review

The pandemic outbreak of COVID-19 has posed a threat to public health globally, and rapid and accurate identification of the viruses is crucial for controlling COVID-19. In recent years, nanomaterial-based electrochemical sensing techniques hold immense potential for molecular diagnosis with high sensitivity and specificity. In this review, we briefly introduced the structural characteristics and routine detection methods of SARS-CoV-2, then summarized the associated properties and mechanisms of the electrochemical biosensing methods. On the above basis, the research progress of electrochemical biosensors based on gold nanomaterials, oxide nanomaterials, carbon-based nanomaterials and other nanomaterials for rapid and accurate detection of virus were reviewed. Finally, the future applications of nanomaterial-based biosensors for biomolecular diagnostics were pointed out. © 2023 Science Press. All rights reserved.

Advanced Nanomaterials-Based Electrochemical Biosensors for Catecholamines Detection: Challenges and Trends.

Catecholamines, including dopamine, epinephrine, and norepinephrine, are considered one of the most crucial subgroups of neurotransmitters in the central nervous system (CNS), in which they act at the brain's highest levels of mental function and play key roles in neurological disorders. Accordingly, the analysis of such catecholamines in biological samples has shown a great interest in clinical and pharmaceutical importance toward the early diagnosis of neurological diseases such as Epilepsy, Parkinson, and Alzheimer diseases. As promising routes for the real-time monitoring of catecholamine neurotransmitters, optical and electrochemical biosensors have been widely adopted and perceived as a dramatically accelerating development in the last decade. Therefore, this review aims to provide a comprehensive overview on the recent advances and main challenges in catecholamines biosensors. Particular emphasis is given to electrochemical biosensors, reviewing their sensing mechanism and the unique characteristics brought by the emergence of nanotechnology. Based on specific biosensors' performance metrics, multiple perspectives on the therapeutic use of nanomaterial for catecholamines analysis and future development trends are also summarized.

An electrochemical PNA-based sensor for the detection of the SARS-CoV-2 RdRP by using surface-initiated-reversible-addition-fragmentation-chain-transfer polymerization technique.

Coronavirus disease 2019 is one of the global health problems. Herein, a highly sensitive electrochemical biosensor has been designed to detect the RNA-dependent RNA polymerase (RdRP) of the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) (SARS-CoV-2 RdRP). Herein, the surface-initiated reversible-addition-fragmentation-chain-transfer polymerization was used to amplify the electrochemical signal. To do that, the thiol-terminated peptide nucleic acid (PNA) probes were first immobilized on the surface of a screen-printed electrode modified with reduced graphene oxide-gold nanocomposite and then the fixed concentration of the SARS-CoV-2 RdRP was added to the electrode surface to interact with PNA probes. Subsequently, the Zr 4+ ions were added to interact with the phosphate groups of the SARS-CoV-2 RdRP. It allowed us to polymerase the ferrocenylmethyl methacrylate (FcMMA) and 4-cyano-4-(phenylcarbonothioylthio)-pentanoic acid on the SARS-CoV-2 RdRP chain. Since the poly-FcMMA has an electrochemical signal, the response of the PNA-based sensor to SARS-CoV-2 RdRP was increased in the range of 5-500 aM. The limit of detection was calculated to be 0.8 aM which is lower than the previous sensor for SARS-CoV-2 RdRP detection. The proposed PNA-based sensor showed high selectivity to the SARS-CoV-2 RdRP in the presence of the gene fragments of influenza A and Middle East respiratory syndrome coronavirus.

Electrochemical detection of Zika and Dengue infections using a single chip.

Zika and Dengue are infectious diseases caused by flaviviruses and transmitted by Aedes mosquitoes. Although symptoms are usually mild, complications such as dengue hemorrhagic fever and microcephaly in newborns -after the pregnant woman becomes infected with the Zika virus-have emerged as a global public health concern. The co-circulation of Zika and Dengue viruses and the overlapping of their symptoms represent a challenge for the accurate diagnosis. A single test for the point-of-care detection of both diseases is crucial. Here we report a single chip that distinguishes between Zika and Dengue infections using the non-structural protein 1 (NS1) as biomarkers. A novel multiplex electrochemical device containing four independent working electrodes was developed. Zika and Dengue biosensors were fabricated separately on different working electrodes. Selectivity tests showed that the two biosensors can distinguish not only the NS1 proteins from Zika and Dengue but also the spike proteins present in the SARS-CoV-2. This is especially relevant as patients with COVID-19 may have symptoms similar to Zika and Dengue. The gold surface was modified with cysteamine and antibodies against the NS1 proteins. Both biosensors detected their respective biomarkers at clinically relevant concentrations and presented a good linear relationship between the percentage change in impedance and the logarithm of the NS1 concentration (R2 = 0.990 for Dengue and R2 = 0.995 for Zika). Upon combining a simple sample preparation with a portable detection method, our disposable multiplex device offers a point-of-care diagnostic test for Zika and Dengue using a single chip. Additionally, two other biosensors can be added to the chip, providing a platform for viral detection.

Functional Nanomaterials for Electrochemical SRAS-CoV-2 Biosensors: a Review

The pandemic outbreak of COVID-19 has posed a threat to public health globally, and rapid and accurate identification of the viruses is crucial for controlling COVID-19. In recent years, nanomaterial-based electrochemical sensing techniques hold immense potential for molecular diagnosis with high sensitivity and specificity. In this review, we briefly introduced the structural characteristics and routine detection methods of SARS-CoV-2, then summarized the associated properties and mechanisms of the electrochemical biosensing methods. On the above basis, the research progress of electrochemical biosensors based on gold nanomaterials, oxide nanomaterials, carbon-based nanomaterials and other nanomaterials for rapid and accurate detection of virus were reviewed. Finally, the future applications of nanomaterial-based biosensors for biomolecular diagnostics were pointed out.

Electrochemical Biosensor for the Determination of Specific Antibodies against SARS-CoV-2 Spike Protein.

In this article, we report the development of an electrochemical biosensor for the determination of the SARS-CoV-2 spike protein (rS). A gold disc electrode was electrochemically modified to form the nanocrystalline gold structure on the surface. Then, it was further altered by a self-assembling monolayer based on a mixture of two alkane thiols: 11-mercaptoundecanoic acid (11-MUA) and 6-mercapto-1-hexanol (6-MCOH) (SAMmix). After activating carboxyl groups using a N-(3-dimethylaminopropyl)-N'-ethyl-carbodiimide hydrochloride and N-hydroxysuccinimide mixture, the rS protein was covalently immobilized on the top of the SAMmix. This electrode was used to design an electrochemical sensor suitable for determining antibodies against the SARS-CoV-2 rS protein (anti-rS). We assessed the association between the immobilized rS protein and the anti-rS antibody present in the blood serum of a SARS-CoV-2 infected person using three electrochemical methods: cyclic voltammetry, differential pulse voltammetry, and potential pulsed amperometry. The results demonstrated that differential pulse voltammetry and potential pulsed amperometry measurements displayed similar sensitivity. In contrast, the measurements performed by cyclic voltammetry suggest that this method is the most sensitive out of the three methods applied in this research.

The critical experimental aspects for developing pathogen electrochemical biosensors: A lesson during the COVID-19 pandemic

Though the bitter global pandemic posed a severe public health threat, it set an unprecedented stage for different research teams to present various technologies for detecting SARS-CoV-2, providing a rare and hard-won lesson for one to comprehensively survey the core experimental aspects in developing pathogens electrochemical biosensors. Apart from collecting all the published biosensor studies, we focused on the effects and consequences of using different receptors, such as antibodies, aptamers, ACE 2, and MIPs, which are one of the core topics of developing a pathogen biosensor. In addition, we tried to find an appropriate and distinctive application scenario (e.g., wastewater-based epidemiology) to maximize the advantages of using electrochemical biosensors to detect pathogens. Based on the enormous amount of information from those published studies, features that fit and favor wastewater pathogen detection can be picked up and integrated into a specific strategy to perform quantitative measurements in wastewater samples. [Display omitted] • Evaluate the effects of different receptors in SARS-CoV-2 electrochemical biosensors. • Dig deep into the rationale why different studies chose specific detection strategies. • Point out the importance of finding appropriate and distinctive application scenarios. • Propose the WBE to maximize the advantages of electrochemical pathogen biosensors. [ FROM AUTHOR]

Detection of COVID-19-related biomarkers by electrochemical biosensors and potential for diagnosis, prognosis, and prediction of the course of the disease in the context of personalized medicine.

As a more efficient and effective way to address disease diagnosis and intervention, cutting-edge technologies, devices, therapeutic approaches, and practices have emerged within the personalized medicine concept depending on the particular patient's biology and the molecular basis of the disease. Personalized medicine is expected to play a pivotal role in assessing disease risk or predicting response to treatment, understanding a person's health status, and, therefore, health care decision-making. This work discusses electrochemical biosensors for monitoring multiparametric biomarkers at different molecular levels and their potential to elucidate the health status of an individual in a personalized manner. In particular, and as an illustration, we discuss several aspects of the infection produced by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) as a current health care concern worldwide. This includes SARS-CoV-2 structure, mechanism of infection, biomarkers, and electrochemical biosensors most commonly explored for diagnostics, prognostics, and potentially assessing the risk of complications in patients in the context of personalized medicine. Finally, some concluding remarks and perspectives hint at the use of electrochemical biosensors in the frame of other cutting-edge converging/emerging technologies toward the inauguration of a new paradigm of personalized medicine.

Customization of aptamer to develop CRISPR/Cas12a-derived ultrasensitive biosensor.

The CRISPR/Cas systems have provided wide biosensing applications. Particularly, the aptamer-involved CRISPR/Cas sensor system powerfully expanded to non-nucleic-acid targets. However, tailoring the sequence of the aptamer to explore the relationship between affinity and the activation of CRISPR/Cas12a trans-cleavage activity has not been reported yet. Herein, we developed a series of new aptamers toward the spike protein 1(S1) of SARS-CoV-2. Surface plasmon resonance measurements showed that the affinity of these aptamers to S1 was at the nM level. Subsequently, a "SET" effect (Sequence Essential Trans-cleavage activity) is discovered for the activation of CRISPR/Cas12a trans-cleavage activity. That is, an aptamer, as the activator, sequence needs to be tailored to activate CRISPR/Cas12a efficiently. A balance should be reached between affinity and activation ability. On the one hand, high affinity ensures target recognition performance, and on the other hand, activation can achieve adequate amplification and output of recognition signals. The optimized sequence (with 27 nucleotides, for short 27-nt) not only recognizes the target with a high affinity and specificity but also can trigger the CRISPR/Cas12a trans-cleavage activity efficiently, showing an excellent detection performance in electrochemical biosensors. The detection limit for SARS-CoV-2 S1 can be low at 1.5 pg mL-1. The new CRISPR/Cas12a-derived aptasensor also displays a remarkable ability to detect Beta, Delta, and Omicron variants but is selective toward other kinds of proteins. Above all, it is robust for point-of-care testing (POCT) in complex biological fluids, such as saliva, urine, and serum, and provides a universal and scalable detecting platform. Our results provide new insights into aptamer development and a different strategy for COVID-19 antigen detection and biosensor development.

Recent advances in electrochemical biosensors – A brief review

A sensitive biological system and a detector system with appropriate transducers for obtaining the output signals make up a biosensor. These devices have a wide range of uses, including disease screening, the detection of environmental pollutants, agriculture, and routine medical examinations. The product's selectivity, sensitivity, stability, and lower production costs will all be critical factors in its widespread commercialisation. Recently, scientists have tackled the issue of developing a nano biosensor with a high degree of sensitivity and selectivity for the recognition of biomarkers of immune responses and cancer. The fact that electrochemical nano biosensors may successfully be employed for detecting medications in addition to pathogen biomarkers has significantly altered the covid picture in our favour. During the worldwide epidemic, a number of cutting-edge SARS - CoV-2 biosensors with portable, smartphone-connected instant detection devices were helpful. This article provides a concise summary of the underlying working principle, generations of developments and numerous detection methods employed by electrochemical biosensors.The difficulties, knowledge gaps, and possible solutions in the field of electrochemical biosensors are all discussed in the context of this categorisation.This novel overview also sheds light on the different kinds of electrochemical biosensors and the tasks they perform, as well as recent developments in the "smart biosensor” industry with potential future directions and challenges for this growing subject.