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A label-free electrochemical strategy is proposed combining equivalent substitution effect with AuNPs-assisted signal amplification. According to the differences of S1 protein in various infectious bronchitis virus (IBV) strains, a target DNA sequence that can specifically recognize H120 RNA forming a DNA-RNA hybridized double-strand structure has been designed. Then, the residual single-stranded target DNA is hydrolyzed by S1 nuclease. Therefore, the content of target DNA becomes equal to the content of virus RNA. After equivalent coronavirus, the target DNA is separated from DNA-RNA hybridized double strand by heating, which can partly hybridize with probe 2 modified on the electrode surface and probe 1 on AuNPs' surface. Thus, AuNPs are pulled to the surface of the electrode and the abundant DNA on AuNPs' surface could adsorb a large amount of hexaammineruthenium (III) chloride (RuHex) molecules, which produce a remarkably amplified electrochemical response. The voltammetric signal of RuHex with a peak near - 0.28 V vs. Ag/AgCl is used as the signal output. The proposed method shows a detection range of 1.56e -9 to 1.56e -6  μM with the detection limit of 2.96e -10  μM for IBV H120 strain selective quantification detection, exhibiting good accuracy, stability, and simplicity, which shows a great potential for IBV detection in vaccine research and avian infectious bronchitis diagnosis. Graphical abstract.

The poly[(9,9-dioctylfuorenyl-2,7-diyl)-alt-co-(1,4-benzo-{2,1',3}-thiadiazole)] (PFBT) was carboxyl-functionalized to prepare polymer dots (C-PFBT Pdots), which served as a self-ECL emitter for producing an extraordinary ECL signal without any exogenous coreactants. The C-PFBT Pdots-modified electrode captured the substrate DNA and further hybridized with a ferrocene (Fc)-labeled DNA. The ECL emission of C-PFBT Pdots was quenched by Fc (a signal off state). After the DNAzyme was added, the DNAzyme-substrate hybrids were formed through hybridizing between DNAzyme and substrate and the Fc-labeled DNA was released. In the presence of target Pb 2+ , the DNAzyme-substrate hybrids could be specifically recognized and cleaved to release the DNAzyme and Pb 2+ . Ultimately, the released DNAzyme would further hybridize with the substrate for producing the DNAzyme-substrate hybrids and then were cleaved by the released Pb 2+ . As a result, the DNA walking machine was generated and the substantial Fc was away from C-PFBT Pdots to obtain a signal on state. Such a strategy achieved a sensitive detection of Pb 2+ and the detection limit was as low as 0.17 pM. Moreover, making this ECL biosensor for an intracellular Pb 2+ detecting, a convincing performance was achieved. The self-ECL emitter C-PFBT Pdots combining with the quencher Fc provided a new strategy and platform for constructing a coreactant-free ECL assay.

In this study, we designed a ZnCdS@ZnS quantum dots (QDs)-based label-free electrochemiluminescence (ECL) immunosensor for sensitive determination of aflatoxin B 1 (AFB 1 ). A Nafion solution assembled abundant QDs on the surface of a Au electrode as ECL signal probes, with specially coupled anti-AFB 1 antibodies as the capturing element. As the reduction reaction between S 2 O 8 2- in the electrolyte and QDs on the electrode led to ECL emission, the decreased ECL signals resulting from target AFB 1 in the samples were recorded for quantification. We evaluated electrochemical impedance spectroscopy and ECL measurements along each step in the construction of the proposed immunosensor. After systematic optimization of crucial parameters, the ECL immunosensor exhibited a good sensitivity, with a low detection limit of 0.01 ng/mL for AFB 1 in a wide concentration range of 0.05-100 ng/mL. Testing with lotus seed samples confirmed the satisfactory selectivity, stability, and reproducibility of the developed ECL immunosensor for rapid, efficient, and sensitive detection of AFB 1 at trace levels in complex matrices. This study provides a powerful and universal analytical platform for a variety of analytes that can be used in broad applications for real-time analysis, such as food and traditional Chinese medicine safety testing, environmental pollution monitoring, and disease diagnostics. Graphical abstract Development of a ZnCdS@ZnS quantum dots based label-free electrochemiluminescence immunosensor for sensitive detection of aflatoxin B 1 in lotus seed.

A titanium dioxide nanoparticle (TiO 2 NP)-mediated resistive biosensor is described for the determination of DNA fragments of Escherichia coli O157:H7 (E. coli O157:H7). The sol-gel method was used to synthesize the TiO 2 NP, and microlithography was applied to fabricate the interdigitated sensor electrodes. Conventional E. coli DNA detections are facing difficulties in long-preparation-and-detection-time (more than 3 days). Hence, electronic biosensor was introduced by measuring the current-voltage (I-V) DNA probe without amplification of DNA fragments. The detection scheme is based on the interaction between the electron flow on the sensor and the introduction of negative charges from DNA probe and target DNA. The biosensor has a sensitivity of 1.67 × 10 13  Ω/M and a wide analytical range. The limit detection is down to 1 × 10 -11  M of DNA. The sensor possesses outstanding repeatability and reproducibility and is cabable to detect DNA within 15 min in a minute-volume sample (1 μL). Graphical abstract Fig. (a) Graphical illustration of electronic biosensor set up and (b) relationship between limit of detection (LOD) and the unaffected poultry samples on E. coli O157:H7.

A facile and highly sensitive biosensor was developed for the determination of hydrogen peroxide (H 2 O 2 ) via electrochemical catalytic reduction of H 2 O 2 by hemoglobin (Hb). Hb was enriched and immobilized simply in a chitosan (Chit) membrane on a magnetic electrode to construct an enzyme-like biosensor. The biosensor catalyzes the electrochemical reduction of H 2 O 2 under an external magnetic field. The response improved roughly twice as Hb was adsorbed by Chit in an alkaline medium. The response of the biosensor under the magnetic field increased by 16% owing to the paramagnetism of Hb. The effect of pH values on Hb adsorption by Chit, as well as the effect of an external magnetic field on Hb configuration were investigated by UV-vis spectroscopy. The reduction peak current has linear and log-linear relationships with H 2 O 2 concentration in the range of 5-250 μmol∙L -1 and 0.01-1 μmol∙L -1 , respectively. The detection limit was 0.003 μmol∙L -1 , with a good sensitivity of 0.227 μA∙μM -1 ∙cm -2 . The biosensor was successfully applied to the determination of H 2 O 2 in milk samples and in disinfectant solutions. Recoveries ranged from 96.3 to 105.4%, and from 95.3 to 107.7%, respectively. Graphical abstractConstruction of the biosensor, and principle of H 2 O 2 determination based on Hb bioelectrocatalysis.

Multiwalled carbon nanotubes coated with cobalt(II) sulfide nanoparticles were prepared and used for immobilization of glucose oxidase (GOx) to obtain an electrochemical glucose biosensor. The nanocomposite was synthesized through an in-situ hydrothermal method and characterized by X-ray diffraction, scanning electron microscopy, transmission electron microscopy, and electrochemical impedance spectroscopy. The results show that the nanocomposite possesses a large specific surface area and apparently enhances the direct electron transfer between GOx and the surface of the electrode, best at a potential near -0.43 V (vs. SCE). The immobilized GOx retains its good bioactivity even at a high surface coverage of 30 pmol cm -2 . Under the optimum conditions. The biosensor exhibits a wide linear range (from 8 μM to 1.5 mM), a high sensitivity (15 mA M -1  cm -2 ), and a 5 μM detection limit (at S/N = 3). The sensor is selective, acceptably repeatable, specific and stable. Graphical abstractMultiwalled carbon nanotubes coated with cobalt(II) sulfide nanoparticles (CoS-MWCNTs) were synthesized through in situ hydrothermal method for the construction of a sensitive electrochemical glucose biosensor.

An electrochemical method is described for the sensitive detection of the activity of the enzyme T4 polynucleotide kinase (PNK) by using a DNA functionalized porphyrinic metal-organic framework (L/(Fe-P)n-MOF). In the presence of PNK, the hairpin oligonucleotide (HP1) becomes phosphorylated, and the trigger is released by lambda exonuclease (λ exo). The trigger DNA hybridizes with hairpin probe (immobilized on the gold electrode) to form a nicking endonuclease cleavage site. Thus, a single-strand capture probe is employed to hybridize with L/(Fe-P)n-MOF. The (Fe-P)n-MOF is a peroxidase mimicking material with high catalytic efficiency. By using this amplification strategy, an electrochemical signal is procured that allows for the determination of T4 PNK in the 1.0 mU·mL -1 to 1.0 U·mL -1 with a detection limit of 0.62 mU·mL -1 . The method is selective and can be used to screen for enzyme inhibitors. Conceivably, the (Fe-P)n-MOF can also be used to detect other analytes via its peroxidase-mimicking activity. Graphical abstract Schematic presentation of T4 polynucleotide kinase (PNK) detection. Two hairpin DNAs (HP) and a porphyrinic metal-organic framework with peroxidase-mimicking activity are used. The detection limit is 0.62 mU mL -1 with enzyme assisted signal amplification. This method is selective and can be used to screen for enzyme inhibitors.

A fluorogenic probe has been developed for determination of telomerase activity using chimeric DNA-templated silver nanoclusters (AgNCs). The formation of AgNCs was investigated before (route A) and after (route B) telomerase elongation reaction. Both routes caused selective quenching of the yellow emission of the AgNCs (best measured at excitation/emission wavelength of 470/557 nm) in telomerase-positive samples. The quenching mechanism was studied using synthetically elongated DNA to mimic the telomerase-catalyzed elongation. The findings show that quenching is due to the formation of parallel G-quadruplexes with a -TTA- loop in the telomerase elongated products. The assay was validated using different cancer cell extracts, with intra- and interassay coefficients of variations of <9.8%. The limits of detection for MCF7, RPMI 2650 and HT29 cell lines are 15, 22 and 39 cells/μL. This represents a distinct improvement over the existing telomeric repeat amplification protocol (TRAP) assay in terms of time, sensitivity and cost. Graphical Abstract A method was developed using chimeric DNA-templated silver nanoclusters to detect telomerase activity directly in cell extracts. The sensitivity of this new method outperforms the traditional TRAP assay, and without the need for amplification.

A small DNA structure, referred to as DNA nanobud (NB), was used for the first time to design a dual-functional nanolabel in order to recognize a particular oligonucleotide sequence, generate and amplify the electrochemical analytical signal. NBs containing numerous repetitive desired sequences were prepared through self-assembly of 8-h rolling circle amplification. Then, redox-active silver ions were loaded onto the NBs by over-night incubation with a solution of AgNO 3 . The incorporation of Ag + into NBs was confirmed by field emission scanning electron microscopy, dynamic light scattering, UV-Vis spectroscopy, zeta potential measurements, and energy-dispersive X-ray spectroscopy. A DNA sandwich complex was created after hybridization of Ag + -NB with target sequence, which was captured by immobilized probe on a gold electrode. Cyclic voltammetry was applied to measure the redox signal of silver ions produced typically at a potential around 0.02 V vs. Ag/AgCl. The label can specifically discriminate fully methylated BMP3 gene from fully unmethylated bisulfate-converted part of the gene. The electrochemical signal produced by DNA sandwich complex of gold/probe/BMP3/Ag + -NB was linear toward BMP3 concentration from 100 pM to 100 nM. The method has a 100 pM BMP3 detection limit. Conceivably, this nanolabel can be designed and modified such that it may also be used to detect other sequences with lower detection limits. Graphical abstract Ag + -NB as a new nanolabel for genosensing was formed by loading Ag + on a spherical DNA nanostructure, nanobud (NB), synthesized by rolling circle amplification process. By using a gold electrode (AuE), Ag + -NB with numerous electroactive cations and binding sites can detect targets and generate amplified electrochemical signals.

The authors describe an electrochemical DNA nanosensor based on the use of single gold nanowire electrodes (AuNWEs). The probe DNA is immobilized on the AuNWE via Au-S bonds that are formed between thiol-terminated DNA and the gold surface. Single AuNWEs were prepared by an improved laser-assisted pulling method and hydrofluoric acid etching. The nanoelectrodes were characterized by cyclic voltammetry and COMSOL simulation. Square wave voltammetry was used to monitor the DNA hybridization event between probe DNA and target DNA by using Methylene Blue (MB) as an intercalator of dsDNA. Under optimal conditions, the peak current for MB (best measured at a potential of -0.2 V vs. Ag/AgCl) increases linearly with the logarithm of the analyte concentration in the 1.0 f. to 10 nM range, with a 0.48 fmM detection limit at an S/N ratio of 3. The assay is highly selective, reproducible and stable. Considering the small overall dimensions and high sensitivity, this nanoelectrode potentially can be applied to in-vivo sensing of DNA inside living cells Graphical abstract Schematic presentation of an electrochemical DNA nanosensor using single gold nanowire electrodes and based on the interaction of thiol-terminated DNA and gold surface. It was used to detect complementary DNA with high selectivity and sensitivity.