Electrochemical Genosensor Research Articles

Advancement in SARS-CoV-2 diagnosis: A new and stable electrochemical biosensor for genomic RNA detection

Coronavirus disease (COVID-19) is caused by infection with the SARS-CoV-2 virus, having already caused more than seven million deaths worldwide. Conventional techniques for SARS-CoV-2 detection have limitations, as high cost, low specificity, and longer analysis time, among others. Biosensors emerge as a necessary alternative to overcome the difficulties of current diagnostics. This paper reports a sensor platform where silver-doped zinc oxide nanomaterial (Ag:ZnONp) was used onto carbon screen-printed electrode and ethidium bromide as indicator for development of a specific electrochemical genosensor for COVID-19. This genosensor demonstrated good linearity between the concentrations of 5.62 × 104–5.62 copies/mL and a detection limit of 5 copies/mL with gRNA in patient’s samples, with a response time within 30 min. Molecular modeling and morphological analysis are in agreement with obtained electrochemical results. Additional techniques such as AFM, SEM, and EIS were conducted to characterize the morphological and electrochemical properties of the biosensor’s surface. The biosensor was also capable of detecting the target presence in spiked samples and demonstrated a stability of 60 days, higher than other similar biosensors for SARS-CoV-2.

Nucleic acid-functionalized nanoscale porous carbon-based electrochemical genosensor for detection of Nocardia spp. in real samples

In this study, a porous carbon derived from a metal-organic framework (PCMOF) as a target-responsive material functionalized with Nocardia particular antisense ssDNA oligonucleotide (ssDNA capture probe) was developed to construct a simple genosensor based on biogatekeeper strategy for sensitive detection of Nocardia in complex biological samples. The PCMOF with suitable pores volume was used to encapsulate electroactive dye methylene blue (MB), and the ssDNA capture probe was used as a gatekeeper to cap PCMOF. Without the presence of Nocardia target, the electrochemical signal of trapped MB was high. Upon adding the target, the hybridization of ssDNA capture probe and target led to the formation of a probe-target double-stranded (dsDNA) structure which dissociated from PCMOF and allowed MB molecules to be released. Therefore, the electrochemical signal of the genosensor decreased. The detection of Nocardia was accomplished by observing variations in the MB peak current intensity in a dose-dependent manner. For this genosensor, a linearity range from 10−18 to 10−7 M for synthetic ssDNA target and 10 to 108 copies/mL for two standard isolates, Nocardia farcinica PTCC 1309 and Nocardia brasiliensis ATCC 19296 as well as for clinical isolates (identified as Nocardia otitidiscaviarum) was observed, respectively. The detection limit (DL) values were 0.54 aM for synthetic ssDNA target and 5, 7, and 4 copies/mL for N. farcinica, N. brasiliensis, and N. otitidiscaviarum, respectively. This genosensor was also characterized by good specificity, reproducibility, and stability.

Development of the BAT-26 mutation-based electrochemical genosensor for identifying microsatellite instability in relationship to cancer

Screening for microsatellite instability is one of the biggest challenges in the cancer diagnosis due to its association with different types of cancer development. In this sense, the development of fast and highly sensitive analytical devices for early the cancer detection is a high priority. In the present work, a highly sensitive electrochemistry genosensor is developed and described for BAT-26 mutation detection using: differential pulse voltammetry (DPV), and cyclic voltametric (CV) supported by scanning tunnel microscopy (STM). The redox reaction of Fe(CN)63−/4– was monitored to probe the presence DNA target. The results demonstrate that modifying the screen-printed gold electrode surface (SPGE) 250AT with a combination of 11-mercaptoundecanoic acid (MUA) and a single-stranded DNA oligonucleotide successfully altered the electrode's properties, particularly its affinity for detecting the BAT-26 mutation under physiological conditions. The genosensor exhibited a wide linear range (100 aM to 1 μM) and achieved a low detection limit of 71.74 aM under optimal parameters. These characteristics, coupled with its high sensitivity and rapid analysis time, make it a promising candidate for development into a portable microsatellite instability diagnostic kit.

An ultrasensitive genosensor for detection of toxigenic and non-toxigenic Clostridioides difficile based on a conserved sequence in surface layer protein coding gene

Clostridioides difficile (C. difficile) is the most common agent of antibiotic-associated diarrhea, leading to intestinal infection through the secretion of two major toxins. Not all strains of this bacterium are toxigenic, but some of them cause infection via their accessory virulence factors, such as surface layer protein (SlpA). SlpA is conserved in both toxigenic and non-toxigenic strains of C. difficile. In the present work, an amplification-free electrochemical genosensor was designed for the detection of the slpA gene. A glassy carbon electrode coated with gold nanoparticle-reduced graphene oxide nanocomposite was used as the working electrode, and its surface was modified using a simple thiolated linear oligonucleotide as the bioreceptor. Moreover, the hexaferrocenium tri[hexa(isothiocyanato) iron(III)] trihydroxonium (HxFc) complex was used as an intercalator, and its redox signal was recorded using differential pulse voltammetry. Scan rate studies indicated a quasi-reversible adsorption-controlled process for the HxFc complex. This genosensor showed high sensitivity with a limit of detection of 0.2 fM, a linear response range of 0.46–1900 fM, and a satisfactory specificity toward the synthetic slpA target gene. Also, the genosensor indicated responses in the mentioned linear range toward the genome extracted from either toxigenic or non-toxigenic strains of C. difficile.

Electrochemical genosensor based on RNA-responsive human telomeric G-quadruplex DNA: A proof-of-concept with SARS-CoV-2 RNA

In this report, a facile and label-free electrochemical RNA biosensor is developed by exploiting methylene blue (MB) as an electroactive positive ligand of G-quadruplex. The electrochemical response mechanism of the nucleic acid assay was based on the change in differential pulse voltammetry (DPV) signal of adsorbed MB on the immobilized human telomeric G-quadruplex DNA with a loop that is complementary to the target RNA. Hybridization between synthetic positive control RNA and G-quadruplex DNA probe on the transducer platform rendered a conformational change of G-quadruplex to double-stranded DNA (dsDNA), and increased the redox current of cationic MB π planar ligand at the sensing interface, thereby the electrochemical signal of the MB-adsorbed duplex is proportional to the concentration of target RNA, with SARS-CoV-2 (COVID-19) RNA as the model. Under optimal conditions, the target RNA can be detected in a linear range from 1 zM to 1 μM with a limit of detection (LOD) obtained at 0.59 zM for synthetic target RNA and as low as 1.4 copy number for positive control plasmid. This genosensor exhibited high selectivity towards SARS-CoV-2 RNA over other RNA nucleotides, such as SARS-CoV and MERS-CoV. The electrochemical RNA biosensor showed DPV signal, which was proportional to the 2019-nCoV_N_positive control plasmid from 2 to 200000 copies (R2 = 0.978). A good correlation between the genosensor and qRT-PCR gold standard was attained for the detection of SARS-CoV-2 RNA in terms of viral copy number in clinical samples from upper respiratory specimens.

Measurement of microRNA-106b as a gastric cancer biomarker based on Zn-BTC MOF label-free genosensor

Due to the late detection of stomach cancer, this cancer usually causes high mortality. The development of an electrochemical genosensor to measure microRNA 106b (miR-106b), as a gastric cancer biomarker, is the aim of this effort. In this regard, first, 1,3,5-benzenetricarboxylate (BTC) metal-organic frameworks (Zn-BTC MOF) were self-assembled on the glassy carbon electrode and then the probe (ssDNA) was immobilized on it. The morphology Zn-BTC MOF was characterized by SEM, FT-IR, Raman and X-Ray techniques. Zn-BTC MOF as a biosensor substrate has strong interaction with ssDNA. Quantitative measurement of miR-106b was performed by electrochemical impedance spectroscopy (EIS). To perform this measurement, the difference of the charge transfer resistances (ΔRct) of Nyquist plots of the ssDNA probe modified electrode before and after hybridization with miR-106b was obtained and used as an analytical signal. Using the suggested genosensor, it is possible to measure miR-106b in the concentration range of 1.0 fM to 1.0 μM with a detection limit of 0.65 fM under optimal conditions. Moreover, at the genosensor surface, miR-106b can be detected from a non-complementary and a single base mismatch sequence. Also, the genosensor was used to assess miR-106b in a human serum sample and obtained satisfactory results.

Novel RNA genosensor based on highly stable gold nanoparticles decorated phosphorene nanohybrid with graphene for highly sensitive and low-cost electrochemical detection of coconut cadang-cadang viroid.

Coconut cadang-cadang viroid (CCCVd) is an infectious single-stranded RNA (ssRNA) pathogen, which leads directly to the death of a large number of coconut palm trees and heavy economic loss to coconut farmers. Herein, a novel electrochemical impedance RNA genosensor is presented based on highly stable gold nanoparticles (AuNPs) decorated phosphorene (BP) nanohybrid with graphene (Gr) for highly sensitive, low-cost, and label-free detection of CCCVd. BP-AuNPs are environmentally friendly prepared by ultrasonic-assisted liquid-phase exfoliation of black phosphorus, accompanying direct reduction of chloroauric acid. Gr/BP-AuNPs are facilely prepared by the in situ growth of AuNPs onto the BP surface and its nanohybrid with Gr to improve environmental stability of BP. Gr/BP-AuNP-based RNA genosensor is fabricated by immobilizing the thiol-functionalized single-stranded DNA (ssDNA) oligonucleotide probe onto the surface of Gr/BP-AuNP-modified glassy carbon electrode via gold-thiol interactions, which served as an electrochemical genosensing platform for the label-free impedance detection of CCCVd by hybridization between the functionalized ssDNA probe and the complementary CCCVd ssRNA sequence in a wide linear range from 1.0 × 10-11 to 1.0 × 10-7M with a low limit of detection of 2.8 × 10-12M. This work supplies an experimental support and theoretical direction for the fabrication of RNA biosensors based on graphene-like materials and potential application for a specific diagnosis of plant RNA viral disease in Arecaceae planting industry.

Detection of high-risk HPV 16 genotypes in cervical cancers using isothermal DNA amplification with electrochemical genosensor

Cervical cancer emerges as the third most prevalent types of malignancy among women on a global scale. Cervical cancer is significantly associated with the persistent infection of human papillomavirus (HPV) type 16. The process of diagnosing is crucial in order to prevent the progression of a condition into a malignant state. The early detection of cervical cancer through initial stage screening is of the utmost significance in both the prevention and effective management of this disease. The present detection methodology is dependent on quantitative polymerase chain reaction (qPCR), which necessitates the use of a costly heat cycler instrument. In this study, we report the development of an electrochemical DNA biosensor integrated with an isothermal recombinase polymerase amplification (RPA) reaction for the detection and identification of the high-risk HPV-16 genotype. The electrochemical biosensor exhibited a high degree of specificity and sensitivity, as evidenced by its limit of detection (LOD) of 0.23 copies/μL of HPV-16 DNA. The validity of this electrochemical platform was confirmed through the analysis of 40 cervical tissues samples, and the findings were consistent with those obtained through polymerase chain reaction (PCR) testing. Our straightforward electrochemical detection technology and quick turnaround time at 75 min make the assay suitable for point-of-care testing in low-resource settings.

Magnetic Beads as Versatile Tools for Electrochemical Biosensing Platforms in Point‐of‐Care Testing

AbstractPoint‐of‐care technologies need particularly stringent requirements, including portability, rapidity, affordability, and ease of use, enabling healthcare practitioners to perform reliable analyses in decentralized settings. Such diagnostic tests are usually based on bioassays that allow for the detection of clinically relevant biomarkers, namely proteins, antibodies and nucleic acids. In the context of electrochemical biosensors, the implementation of magnetic beads offers unique advantages stemming from their high surface‐to‐volume ratio, easy manipulation through magnetic fields and versatility in surface chemistries for bio‐conjugation. The present concept article highlights the advantages achieved through the implementation of magnetic beads in electrochemical binding assays, immunosensors and genosensors. These methods mostly exploit magnetic beads for immobilization purposes, allowing for the assembly of bio‐complexes in liquid suspensions under precise temperature and shaking control, as well as simplifying the manipulation steps to enable untrained personnel to perform analyses. In addition, magnetic beads can also take part in the detection mechanism, leading to innovative sensing strategies with great potential for point‐of‐care diagnostic applications.

A label-free sensor based on a carbon nanotube-graphene platform for the detection of non-Hodgkin lymphoma genes

Early diagnosis of non-Hodgkin lymphoma (NHL) is crucial for effective management. We report an ultrasensitive electrochemical genosensor using multi-walled carbon nanotube (MWCNT)-graphene nanocomposite for detection of lymphoma biomarker gene. MWCNTs were dispersed over graphene nanoplatelets to form a conductive nanocomposite platform. The high surface area and π-π stacking ability enabled efficient immobilization of lymphoma probe DNA. Detailed materials characterization was performed using microscopy and spectroscopy techniques. The sensor was fabricated by drop-casting probe-conjugated nanocomposite suspension onto a glassy carbon electrode. The genosensor exhibited a wide dynamic range of 1 fM to 1 nM with a sensitivity of 0.895 kΩ per decade of target concentration. The sensor showed excellent selectivity for fully complementary target sequence over even single base mismatch. Hybridization detection by electrochemical impedance spectroscopy was further validated using differential pulse voltammetry which showed drastic reduction in redox peak current after incubating with target. The nanocomposite genosensor demonstrated good reproducibility with relative standard deviation of 3.72 % among five independently prepared electrodes. Clinical applicability was tested by spiking lymphoma gene biomarker in 10 % serum and recoveries of 93–99 % were obtained. The proposed MWCNT-graphene nanocomposite genosensor provides a simple, specific and ultrasensitive platform for point-of-care diagnosis of NHL.

Ultrasensitive rapid detection of antibiotic resistance genes by electrochemical ratiometric genosensor based on 2D monolayer Ti3C2@AuNPs

As an important emerging pollutant, antibiotic resistance genes (ARGs) monitoring is crucial to protect the ecological environment and public health, but its rapid and accurate detection is still a major challenge. In this study, a new single-labeled dual-signal output ratiometric electrochemical genosensor (E-DNA) was developed for the rapid and highly sensitive detection of ARGs using a synergistic signal amplification strategy of T3C2@Au nanoparticles (T3C2@AuNPs) and isothermal strand displacement polymerase reaction (ISDPR). Specially, two-dimensional monolayer T3C2 nanosheets loaded with uniformly gold nanoparticles were prepared and used as the sensing platform of the E-DNA sensor. Benefiting from excellent conductivity and large specific surface area of Ti3C2@AuNPs, the probe immobilization capacity of the E-DNA sensor is doubled, and electrochemical response signals of the E-DNA sensor were significantly improved. The proposed single-labeled dual-signal output ratiometric sensing strategy exhibits three to six times higher sensitivity for the sul2 gene than the single-signal sensing strategy, which significantly reduces cost meanwhile retaining the advantages of high sensitivity and reliability offered by conventional dual-labeled ratiometric sensors. Coupled with ISDPR amplification technology, the E-DNA sensor has a wider linear range from 10 fM to 10 nM and a limit of detection as low as 2.04 fM (S/N=3). More importantly, the E-DNA sensor demonstrates excellent specificity, good stability and reproducibility for target ARGs detection in real water samples. The proposed new sensing strategy provides a highly sensitive and versatile tool for the rapid and accurate quantitative analysis of various ARGs in environmental water samples.

Cerium-Encapsulated SbIII-SeIV-Templating Polyoxotungstate for Electrochemically Sensing Human Multidrug Resistance Gene Segment.

A tower-like SbIII-SeIV-templating polyoxotungstate [H2N(CH3)2]12Na7H3[Ce0.5/Na0.5(H2O)5]2[SbSe2W21O75]2·50H2O (1) was synthesized, whose skeleton is assembled from two prolonged lacunary Dawson [SbSe2W21O75]13- units and two [Ce0.5/Na0.5(H2O)5]2+ linkers. The uncommon [SbSe2W21O75]13- unit can be viewed as a combination of one [SeW6O21]2- group grafted onto a trivacant Dawson [SbSeW15O54]11- subunit. The conductive composite 1-Au@rGO containing 1, gold nanoparticles, and reduced graphene oxide (rGO) was conveniently prepared, using which the 1-Au@rGO-based electrochemical genosensor was constructed for detecting human multidrug resistance gene segment. This work enriches structural types of dual-heteroatom-inserted polyoxometalates and promotes the application of polyoxometalates in genosensors.

Flexible genosensors based on polypyrrole and graphene quantum dots for PML/RARα fusion gene detection: A study of acute promyelocytic leukemia in children

Acute promyelocytic leukemia (APL) in children is associated with a favorable initial prognosis. However, minimal residual disease (MRD) follow-up remains poorly defined, and relapse cases are concerning due to their recurrent nature. Thus, we report two electrochemical flexible genosensors based on polypyrrole (PPy) and graphene quantum dots (GQDs) for label-free PML-RARα oncogene detection. Atomic force microscopy (AFM), scanning electron microscope (SEM), cyclic voltammetry (CV), and electrochemical impedance spectroscopy (EIS) were used to characterize the technological biosensor development. M7 and APLB oligonucleotide sequences were used as bioreceptors to detect oncogenic segments on chromosomes 15 and 17, respectively. AFM characterization revealed heterogeneous topographical surfaces with maximum height peaks for sensor layers when tested with positive patient samples. APLB/Genosensor exhibited a percentage change in anode peak current (ΔI) of 423 %. M7/Genosensor exhibited a ΔI of 61.44 % for more concentrated cDNA samples. The described behavior is associated with the biospecific recognition of the proposed biosensors. Limits of detection (LOD) of 0.214 pM and 0.677 pM were obtained for APLB/Genosensor and M7/Genosensor, respectively. The limits of quantification (LOQ) of 0.648 pM and 2.05 pM were estimated for APLB/Genosensor and M7/Genosensor, respectively. The genosensors showed reproducibility with a relative standard deviation of 7.12 % for APLB and 1.18 % for M7 and high repeatability (9.89 % for APLB and 1.51 % for M7). In addition, genetic tools could identify the PML-RARα oncogene in purified samples, plasmids, and clinical specimens from pediatric patients diagnosed with APL with high bioanalytical performance. Therefore, biosensors represent a valuable alternative for the clinical diagnosis of APL and monitoring of MRD with an impact on public health.

An Ultrasensitive Voltammetric Genosensor for the Detection of Bacteria Vibrio cholerae in Vegetable and Environmental Water Samples.

In view of the presence of pathogenic Vibrio cholerae (V. cholerae) bacteria in environmental waters, including drinking water, which may pose a potential health risk to humans, an ultrasensitive electrochemical DNA biosensor for rapid detection of V. cholerae DNA in the environmental sample was developed. Silica nanospheres were functionalized with 3-aminopropyltriethoxysilane (APTS) for effective immobilization of the capture probe, and gold nanoparticles were used for acceleration of electron transfer to the electrode surface. The aminated capture probe was immobilized onto the Si-Au nanocomposite-modified carbon screen printed electrode (Si-Au-SPE) via an imine covalent bond with glutaraldehyde (GA), which served as the bifunctional cross-linking agent. The targeted DNA sequence of V. cholerae was monitored via a sandwich DNA hybridization strategy with a pair of DNA probes, which included the capture probe and reporter probe that flanked the complementary DNA (cDNA), and evaluated by differential pulse voltammetry (DPV) in the presence of an anthraquninone redox label. Under optimum sandwich hybridization conditions, the voltammetric genosensor could detect the targeted V. cholerae gene from 1.0 × 10-17-1.0 × 10-7 M cDNA with a limit of detection (LOD) of 1.25 × 10-18 M (i.e., 1.1513 × 10-13 µg/µL) and long-term stability of the DNA biosensor up to 55 days. The electrochemical DNA biosensor was capable of giving a reproducible DPV signal with a relative standard deviation (RSD) of <5.0% (n = 5). Satisfactory recoveries of V. cholerae cDNA concentration from different bacterial strains, river water, and cabbage samples were obtained between 96.5% and 101.6% with the proposed DNA sandwich biosensing procedure. The V. cholerae DNA concentrations determined by the sandwich-type electrochemical genosensor in the environmental samples were correlated to the number of bacterial colonies obtained from standard microbiological procedures (bacterial colony count reference method).

A highly reusable genosensor for late-life depression diagnosis based on microRNA 184 attomolar detection in human plasma.

Late-Life Depression (LLD) is one of the most prevalent psychiatric disorders in elderly, causing significant functional impairments. MicroRNAs are small molecules involved in the post-transcriptional regulation of gene expression. Elderly individuals diagnosed with LLD present down regulation of miR-184 (hsa-miR-184) expression compared to healthy patients. Therefore, this miR-184 can be used as a biomarker to diagnose LLD. Current LLD diagnosis depends primarily on clinical subjective identification, based on symptoms and variable scales. This work introduces a novel and facile approach for the LLD diagnosis based on the development of an electrochemical genosensor for miR-184 detection in plasma, using differential pulse voltammetry (DPV) and electrochemical impedance spectroscopy (EIS). DPV results presented a 2-Fold increase in current value for healthy patients, compared to individuals with LLD when monitoring ethidium bromide oxidation peak. For EIS, a 1.5-fold increase in charge transfer resistance for healthy elderly subjects was observed in comparison with depressed patients. In addition, the analytical performance of the biosensor was evaluated using DPV, obtaining a linear response ranging from 10-9molL-1 to 10-17molL-1 of miR-184 in plasma and a detection limit of 10 atomoles L-1. The biosensor presented reusability, selectivity and stability, the current response remained 72% up to 50 days of storage. Thus, the genosensor proved to be efficient in the diagnosis of LLD, as well as the accurate quantification of miR-184 in real plasma samples of healthy and depressed patients.

Recycled additive manufacturing feedstocks with carboxylated multi-walled carbon nanotubes toward the detection of yellow fever virus cDNA

Recycled additive manufacturing sensing platforms are fabricated with carboxylated multi-walled carbon nanotubes (COOH-MWCNT) with exhibit enhanced electrochemical biosensor performance allowing for the enhanced direct coupling of the biorecognition element to the COOH-MWCNT for the preparation of an electrochemical genosensor for the detection of yellow fever virus cDNA. Bespoke additive manufacturing filaments was produced using recycled poly(lactic acid) (rPLA, 65 wt%), polyethylene succinate (PES, 10 wt%), carbon black (CB, 15 wt%), and COOH-MWCNT (10 wt%) which exhibits enhanced electrochemical performance over that of commercial filament. A bespoke all-in-one additive manufactured electroanalytical cell is proposed, with the working, reference and counter electrodes in addition to a modification rim that allows for the facile production of biosensors through the application of droplets. The genosensor was applied to the detection of yellow fever Virus cDNA using anodic square wave voltammetry; a linear dynamic range (LDR) of 0.5–15 µM with an R2 of 0.9995, sensitivity of 177 ± 2 µA µM−1, limit of detection (LOD) of 0.138 µM, and limit of quantification (LOQ) of 0.859 µM were obtained. This work highlights how bespoke additive manufacturing filament production can enhance biosensing platforms, whilst using recycled feedstock to improve end-product sustainability.

A sensitive electrochemical genosensor for measuring gastric cancer biomarker, microRNA-106b, based on asparagine polymerization on carbon quantum dots modified electrode

The aim of this work is to develop an electrochemical genosensor to measure microRNA 106b (miR-106b) as a gastric cancer biomarker. To construct the miR-106b genosensor, asparagine (As) was electropolymerized on the surface of carbon quantum dots modified glassy carbon electrode (CQDs/GCE). In the next step, ssDNA was immobilized on the modified electrode surface (As/CQDs/GCE) and then hybridized with miR-106b. Finally, differential pulse voltammograms of the accumulated chlorogenic acid (CGA), as an electroactive label, on the surface of immobilized ssDNA was recorded before (ssDNA/As/CQDs/GCE) and after hybridization with miR-106b (miR-106b/BSA/ssDNA/As/CQDs/GCE). The difference in the peak current of these two voltammograms was used as an analytical signal to measure of miR-106b. Using this genosensor, a linear concentration range from 1.0 fM to 1.0 µM and a detection limit of 0.39 fM were obtained for the measurement of miR-106b. The obtained data indicate that it is possible to recognize miR-106 from a non-complementary and a single-base mismatch sequence using the genosensor. In addition, the measurement of miR-106b in a human serum sample was successfully performed using the genosensor.

Electrochemical Detection of ompA Gene of C. sakazakii Based on Glucose-Oxidase-Mimicking Nanotags of Gold-Nanoparticles-Doped Copper Metal-organic Frameworks.

The present work developed an electrochemical genosensor for the detection of virulence outer membrane protein A (ompA, tDNA) gene of Cronobacter sakazakii (C. sakazakii) by exploiting the excellent glucose-oxidase-mimicking activity of copper Metal-organic frameworks (Cu-MOF) doped with gold nanoparticle (AuNPs). The signal nanotags of signal probes (sDNA) that biofunctionalized AuNPs@Cu-MOF (sDNA-AuNPs@Cu-MOF) were designed using an Au-S bond. The biosensor was prepared by immobilization capture probes (cDNA) onto an electrodeposited AuNPs-modified glassy carbon electrode (GCE). AuNPs@Cu-MOF was introduced onto the surface of the GCE via a hybridization reaction between cDNA and tDNA, as well as tDNA and sDNA. Due to the enhanced oxidase-mimicking activity of AuNPs@Cu-MOF to glucose, the biosensor gave a linear range of 1.0 × 10-15 to 1.0 × 10-9 mol L-1 to tDNA with a detection limit (LOD) of 0.42 fmol L-1 under optimized conditions using differential pulse voltammetry measurement (DPV). It can be applied in the direct detection of ompA gene segments in total DNA extracts from C. sakazakii with a broad linear range of 5.4-5.4 × 105 CFU mL-1 and a LOD of 0.35 CFU mL-1. The biosensor showed good selectivity, fabricating reproducibility and storage stability, and can be used for the detection of ompA gene segments in real samples with recovery between 87.5% and 107.3%.

Bio-inspired polynorepinephrine based nanocoatings for reduced graphene oxide/gold nanoparticles composite for high-performance biosensing of Mycobacterium tuberculosis

Polydopamine (PDA) has established itself as a promising grafting and coating material, particularly for functional group-deprived electrochemically active nanomaterials such as graphene, MXene, CNT, metal nanoparticles, and so on, and has proven its extensive applicability in the design and development of electrochemical biosensor devices. However, polynorepinephrine (PNE), a sister compound of PDA, having additional –OH groups and greater coating uniformity and biocompatibility, has never been studied in the field of biosensors. Herein, we investigated PNE as a coating material for reduced graphene oxide (RGO) and gold nanoparticles (Au) in order to build an electrochemical genosensor for Mycobacterium tuberculosis (MTB) detection. Biotin-Avidin chemistry was used to covalently immobilize probe DNA (ssDNA) specific to MTB to the nanocomposite surface on glassy carbon electrode (GCE) in order to construct biosensing electrodes. The formation of RGO/PNE and RGO/PNE/Au nanocomposite as well as the immobilization of ssDNA onto the bioelectrodes are both corroborated by UV–Visible, Raman, and XRD studies with FE-SEM and HR-TEM analysis. The electrochemical studies performed using cyclic voltammetry (CV) and linear sweep voltammetry (LSV) showed the significant enhancement in charge transfer kinetics of RGO/PNE/GCE and RGO/PNE/Au/GCE electrode compared to GO/GCE electrode. The biosensing investigations performed using ssDNA/avidin/RGO/PNE/Au/GCE bioelectrode showed high sensitivity (2.3 × 10−3 mA μM−1), low detection limit (0.1 × 10−7 μM), broad detection range (0.1 × 10−2 to 0.1 × 10−7 μM) with good selectivity and low response time (5 s) of the developed sensor. In comparison to the analogous RGO/PDA/Au material system, RGO/PNE/Au demonstrated increased enzyme loading, improved electrochemical responsiveness, and superior biosensing performance.

Reduced Graphene Oxide-Polydopamine-Gold Nanoparticles: A Ternary Nanocomposite-Based Electrochemical Genosensor for Rapid and Early Mycobacterium tuberculosis Detection.

Tuberculosis (TB) has been a devastating human illness for thousands of years. According to the WHO, around 10.4 million new cases of tuberculosis are identified every year, with 1.8 million deaths. To reduce these statistics and the mortality rate, an early and accurate TB diagnosis is essential. This study offers a highly sensitive and selective electrochemical biosensor for Mycobacterium tuberculosis (MTB) detection based on a ternary nanocomposite of reduced graphene oxide, polydopamine, and gold nanoparticles (rGO-PDA-AuNP). Avidin-biotin coupling was used to bind the MTB probe DNA onto the rGO-PDA-AuNP modified glassy carbon electrode (ssDNA/avidin/rGO-PDA-AuNP). UV-Visible, Raman, XRD, and TEM were used to evaluate the structural and morphological characteristics of rGO-PDA-AuNP. Furthermore, DNA immobilization is validated using FESEM and FT-IR techniques. The modified electrodes were electrochemically analyzed using cyclic voltammetry (CV) and linear sweep voltammetry (LSV), and the results indicate that the produced electrode can detect target DNA up to 0.1 × 10-7 mM with 2.12 × 10-3 mA µM-1 sensitivity and a response time of 5 s. The constructed genosensor displayed high sensitivity and stability, and it also provides a unique strategy for diagnosing MTB at an early stage. Furthermore, our rGO-PDA-AuNP/GCE-based electrochemical platform has broad potential for creating biosensor systems for detecting various infectious pathogens and therapeutically significant biomarkers.