Dual Signal Research Articles

A hemin/rGO/MWCNT nanocomposite-based dual signal electrochemical aptasensor for sensitive detection of NSE.

Neuron-specific enolase (NSE), a tumor marker of small cell lung cancer (SCLC), has high application value in the early diagnosis of SCLC. In this study, a dual signal electrochemical aptasensor for NSE was constructed based on hemin/reduced graphene oxide/multi-walled carbon nanotube (H-rGO-MWCNT) nanocomposites. Hemin played a dual role, functioning not only as an in situ electrochemical probe but also exhibiting excellent peroxidase-like properties, effectively catalyzing the electroreduction of H2O2. Reduced graphene oxide and multi-walled carbon nanotubes exhibited excellent conductivity. Through their binding with hemin, the nanocomposites achieved a larger specific surface area, providing numerous active sites for capturing the NSE aptamer. In the presence of NSE, the specific adsorption between the antigen and the aptamer formed a stable antigen-aptamer structure, which inhibited the performance of hemin, resulting in the weakening of the electrochemical signals of hemin and H2O2. Leveraging these characteristics, the sensitive and cost-effective dual-signal electrochemical aptasensor has been fabricated for the detection of NSE. One signal corresponded to differential pulse voltammetry (DPV) of hemin, while the other signal was derived from chronoamperometry, capturing the catalytic reduction of H2O2. The linear ranges for NSE were 1 pg mL-1 to 1 μg mL-1 and 100 pg mL-1 to 100 ng mL-1 with the limit of detection (LOD) of 0.21 pg mL-1 and 11.22 pg mL-1 by DPV and chronoamperometry, respectively. In addition, this aptasensor exhibited good reproducibility, stability and specificity. The recovery of NSE in human blood serum samples was from 89% to 131%. It provided a promising strategy for the detection of NSE in clinical diagnostics.

Label-free ratiometric fluorescence detection of Pb2+via structure-specific fluorescent dyes and dual signal amplification.

Lead ions (Pb2+) are a widely distributed and highly toxic heavy metal pollutant, which seriously threatens the environment, economy and human safety. Here, a label-free ratiometric fluorescent biosensor was constructed for Pb2+ detection using DNAzyme-driven target cycling and exonuclease III (Exo III)-mediated DNA cycling as a dual signal amplification strategy. The SYBR Green I (SGI) and N-methyl mesoporphyrin IX (NMM) used in this study are characterized by low cost, storage resistance, and short preparation time compared with conventional signaling probes labeled with fluorescent groups. Unlike the single-emission fluorescence strategy, monitoring the fluorescence intensity ratio of SGI and NMM can effectively reduce external interference to achieve accurate detection of Pb2+. DNAzyme structures on the surface of magnetic beads (MBs) can recognize Pb2+ and activate the target circulatory system to cleave single-stranded DNA (ssDNA). The ssDNA further initiated the Exo III-assisted DNA circulatory system to digest double-stranded DNA (dsDNA) and release guanine-rich G1. Finally, the fluorescence signals of SGI and NMM were weakened and enhanced, respectively. The sensing strategy achieved a wide linear range from 0.5 to 500 nM and a low limit of detection (LOD) of 26.4 pM. Furthermore, its anti-interference ability and potential applicability for Pb2+ detection in actual samples were verified. This work ingeniously combines the dual signal amplification strategy with the ratiometric sensing strategy constructed by structure-specific fluorescent dyes, which provides a promising method for constructing sensitive and accurate fluorescent biosensors.

Plasmonic and nanozyme dual-channel-based logic judgment for enhancing gold nanoparticle based colorimetric Hg2+ ion sensing performance.

An AND logic gate-based Hg2+ ion colorimetric assay was constructed using the plasmonic and nanozyme dual signal channels of gold nanoparticles (AuNPs). This assay increased the judgment criteria for the identification of Hg2+ ions and effectively improved the accuracy of Hg2+ ion detection.

Simple and direct electrochemical detection of rosmarinic acid in food samples based on nanochannel modified carbon electrode.

The detection of rosmarinic acid (Ros A) in food samples holds major significance. Simple and convenient electrochemical detection of Ros A with high performance remains a challenge. In this work, a nanochannel array-modified carbon electrode was constructed using a simple and convenient approach to achieve highly sensitive electrochemical detection of Ros A in food samples. Through simple electrochemical pre-activation of a glassy carbon electrode (GCE), oxygen-containing functional groups were introduced on the electrode surface (p-GCE). Vertically-ordered mesoporous silica film (VMSF) was stably grown on p-GCE through electrochemical-assisted self-assembly (EASA) without the introduction of another adhesive layer (VMSF/p-GCE). Transmission electron microscopy (TEM) characterization demonstrated the highly ordered structure of VMSF with a nanochannel diameter around 2.7 nm. Both p-GCE and the nanochannels significantly enhanced the electrochemical signals of Ros A on the electrode, exhibiting dual signal amplification. VMSF/p-GCE demonstrated sensitive detection of Ros A with a linear range of 500 nM to 1 μM and 1 μM to 35 μM. The detection limit (DL) was 26 nM. Combining the good anti-fouling and anti-interference properties of the nanochannels, VMSF/p-GCE can achieve direct electrochemical detection of Ros A in food samples. The sensor can be easily regenerated for repeated use. The simple fabrication, high detection sensitivity and selectivity of the sensor make it a new strategy for rapid preparation of high-performance electrochemical sensors.

Detection of free DNA based on metal-enhanced fluorescence triggered by CRISPR-Cas12a and colorimetric analysis.

The CRISPR-Cas system has been found to be extremely sensitive and there is an urgent demand to extend its potential in bioassays. Herein, we developed a novel nanobiosensor to detect the human papillomavirus 16 genes (HPV-16 DNA), which is triggered by CRISPR-Cas12a to amplify the fluorescence signal by metal-enhanced fluorescence (CAMEF). Along with the changing of the fluorescence signal, the aggregation of the substrate of MEF also leads to a change in the color of the mixture solution, enabling dual signal detection with the fluorescence and the naked eye. Furthermore, the designed CAMEF probe was verified to detect the HPV-16 DNA accurately and reliably in biological samples. Triggered by the CRISPR system, the designed CAMEF probe allows quantitative detection of the HPV-16 DNA in the wide range of 10-500 pM. Owing to the MEF, the fluorescence signal of the CAMEF probe was significantly amplified with the detection limit as low as 1 pM. Besides, we can determine the concentration of HPV-16 DNA simply by the naked eye, which also drastically reduces the possibility of false-positive signals. Theoretically, the target ssDNA could be any strand of DNA obtained by designing the crRNA sequence in the CRISPR-Cas system. We believe that the designed CAMEF sensor can present a reliable approach for the accurate detection of low amounts of target ssDNA in complex biological samples.

Biomimetic photonic crystal double-network hydrogel for visual and electrical dual signal bluetooth-enabled wearable sensor

The photonic crystal wearable sensor which has visual and electrical dual signal outputs is successfully developed.

Synthesis of a benzothiazole-based structure as a selective colorimetric-fluorogenic cyanate chemosensor

Abstract In this study, a new heterocyclic compound incorporating a benzothiazole moiety was specifically designed for the detection of cyanate anions, employing a hydrogen bonding mechanism. Through strategic integration of triazine and phenylenediamine cyclic groups into the compound’s structure, intramolecular hydrogen bonding interactions between the donor and acceptor sites were enhanced, leading to exceptional sensitivity towards cyanate anions. Utilizing the amino-type excited-state intramolecular proton transfer phenomenon, this new compound exhibited dual signals and achieved a significant Stokes’ shift via proton transfer, coupled with aggregation-induced emission properties. This unique combination resulted in visible color changes and an impressive fluorescence response, offering a promising solution for the sensitive detection of cyanate ions in critical environmental matrices. Cyanate detection at low concentrations by this as-synthesized compound (L1), accompanied by a distinct color change and a gradual fluorescence increase upon incremental cyanate addition demonstrated L1’s selectivity, as confirmed in the presence of various competing anions F−, Cl−, Br−, I−, ClO 3 − {\text{ClO}}_{3}^{-} , ClO 4 − {\text{ClO}}_{4}^{-} , NO 3 − {\text{NO}}_{3}^{-} , BrO 3 − {\text{BrO}}_{3}^{-} , CN− and CNO−. Spectrofluorometric investigations demonstrated that L1 shows significant potential as a selective cyanate anion detection candidate.

Hemin/G-quadruplex and AuNPs-MoS2 based novel dual signal amplification strategy for ultrasensitively sandwich-type electrochemical thrombin aptasensor

In this work, a novel sandwich-type electrochemical aptasensor based on the dual signal amplification strategy of hemin/G-quadruplex and AuNPs-MoS2 was designed and constructed, which realized the highly sensitive and specific detection of thrombin (TB). In this aptasensor, the 15-mer TB-binding aptamer (TBA-1) modified with thiol group was immobilized on the surface of AuNPs modified glassy carbon electrode (AuNPs/GCE) as capturing elements. Another thiol-modified 29-mer TB-binding aptamer (TBA-2) sequence containing G-quadruplex structure for hemin immobilization was designed. The formed hemin/G-quadruplex/TBA-2 sequence was further combined to the AuNPs decorated flower-like molybdenum disulfide (AuNPs-MoS2) composite surface via Au-S bonds, acting the role of reporter probe. In presence of the target TB, the sandwich-type electrochemical aptamer detection system could be formed properly. With the assistance of the dual signal amplification of AuNPs-MoS2 and hemin/G-quadruplex toward H2O2 reduction, the sandwich-type electrochemical aptasensor was successfully constructed for sensitive detection of TB. The results demonstrate that the fabricated aptasensor displays a wide linear range of 1.0 × 10−6 ∼ 10.0 nM with a low detection limit of 0.34 fM. This proposed aptasensor shows potential application in the detection of TB content in real biological samples with high sensitivity, selectivity, and reliability.

Serving two masters – How dual price signals can undermine demand flexibility

Demand-side flexibility is often encouraged via demand response programs, where users are incentivized to adjust their electricity consumption based on price signals in tariffs. The design and implementation of these signals are vital, as they link conditions of the energy system with user behaviour. As a result of energy system separation into stages of generation, many users are required to simultaneously respond to two price signals. While existing research has mainly examined responses to a single signal, this study highlights the complexities when users must react to two signals simultaneously. It presents a case study using two distinct price signals: real-time pricing (RTP) and a demand charge, analysing the implications such dual price signals could have on users. The study reveals the empirical incompatibility of these signals, leading to conflicting user goals. Such interference complicates decision-making for demand flexibility, potentially eroding user trust in retailers and system operators. The study ends with proposing an alternative tariff, resolving the dual price situation, which facilitates user comprehension and decision-making, and thus enhances the potential for demand flexibility.

A colorimetric and fluorescent dual-mode sensor based on bifunctional G-quadruplex-hemin complex for the determination of Pb2+

Due to the threat of lead pollution to health, environmental and food safety, developing simple and fast detection methods is highly required. Whereas, traditional single-mode probe suffers from limited application scenario. In this study, a colorimetric and fluorometric dual-mode probe for Pb2+ determination was constructed by using bifunctional G-quadruplex-hemin complex. In this dual-mode probe, enzyme strand and substrate strand of 8–17 DNAzyme are labeled with G-quadruplex-hemin complex and fluorophore, respectively. In the absence of Pb2+, the self-assembly of enzyme strand and substrate strand inhibits intrinsic mimic peroxidase of G-quadruplex-hemin complex by base-pairing, which also quench the fluorescence via in proximity effect. When the DNAzyme is activated by Pb2+, the quenched fluorescence is restored as well as the inherent peroxidase mimetic activity, leading to dual signal output. Under optimal conditions, this dual-mode probe exhibit a good linear relationship between logarithm of Pb2+ concentration and signal difference within the range from 1.5 nM to 20 nM and 0.5 nM to 10 nM for colorimetric and fluorescence mode, respectively. The detection limits for the corresponding mode were estimated to be 1.29 nM and 0.16 nM, respectively. This dual-mode probe also successfully applied for the spiked river water assay with satisfactory recovery in the range of 93.2 %–115.3 %. This work paves a new way for DNAzyme based dual-mode probe construction.

Dual signal amplification platform based on recombinant polymerase amplification and fluorescence-enhanced nanoparticles for sensitive detection of Salmonella

Salmonella is a common foodborne pathogen, and its detection is essential for managing public food safety. Lateral flow assay (LFA) strips are well suited for the purpose as they are inexpensive and easy to use. However, the sensitive detection of low-concentration targets requires the use of signal amplification strategies. This paper presents a LFA for the ultrasensitive detection of Salmonella based on recombinase polymerase amplification (RPA) and fluorescence-enhanced composite nanospheres dSiO2@CD@SiO2 (dCDS). RPA is capable of rapid (∼20 min) exponential amplification of bacterial DNA and does not require complex instrumentation. Mesoporous silica (dSiO2) has a large specific surface area and radial pore size, making it an excellent structural material for internal assembly. Nanoparticles obtained by loading carbon dots (CDs) had a strong fluorescence signal and stability. We used dCDSs as reporter molecules, which captured a large number of DNA-dCDS complexes on the test line of the LFA. With double signal amplification, the RPA and dCDS-based LFA showed greatly improved sensitivity: The limits of detection for Salmonella in buffer and milk were 3.8 CFU/mL and 36 CFU/mL, respectively. This study provides an inexpensive strategy for the sensitive detection of pathogenic bacteria.

Effects of hyperglycemia on the TGF-β pathway in trabecular meshwork cells

BackgroundHyperglycemia, which can lead to apoptosis, hypertrophy, fibrosis, and induces hyperinflammation in diabetic vascular complications due to oxidative stress. In order to elucidate the potential dual roles and regulatory signal transduction of TGF-β1 and TGF-β2 in human trabecular meshwork cells (HTMCs), we established an oxidative cell model in HTMCs using 5.5, 25, 50, and 100 mM d-glucose-supplemented media and characterized the TGF-β-related oxidative stress pathway. MethodsFurther analysis was conducted to investigate oxidative damage and protein alterations in the HTMC caused by the signal transduction. This was done through a series of qualitative cell function studies, such as cell viability/apoptosis analysis, intracellular reactive oxygen species (ROS) detection, analysis of calcium release concentration, immunoblot analysis to detect the related protein expression alteration, and analysis of cell fibrosis to study the effect of different severities of hyperglycemia. Also, we illustrated the role of TGF-β1/2 in oxidative stress-induced injury by shRNA-mediated knockdown or stimulation with recombinant human TGF-β1 protein (rhTGF-β1). ResultsResults from the protein expression analysis showed that p-JNK, p-p38, p-AKT, and related SMAD family members were upregulated in HTMCs under hyperglycemia. In the cell functional assays, HTMCs treated with rhTGFβ-1 (1 ng/mL) under hyperglycemic conditions showed higher proliferation rates and lower ROS and calcium levels. ConclusionsTo summarize, mechanistic analyses in HTMCs showed that hyperglycemia-induced oxidative stress activated TGF-β1 along with its associated pathway. General Significance: While at low concentrations, TGF-β1 protects cells from antioxidation, whereas at high concentrations, it accumulates in the extracellular matrix, causing further HTMC dysfunction.

Calibrating nonlinearity coefficients of a nano-g accelerometer by dual-frequency excitation on a shaker

Nano-g accelerometers are utilized extensively in high precision applications, such as gravity measurements on moving bases, which often face strong motion disturbances. Accurately-calibrated coefficients are crucial for suppressing their high-order error effects, but challenging to measure using conventional calibration methods. This paper presents an investigation into calibration of the nonlinearity coefficient of nano-g accelerometers on a horizontal shaker driven by a dual frequency signal. A nonlinear output component occurring at the low difference frequency, with a high signal-to-noise ratio, is utilized. The calibration principle and strategy are investigated, along with an analysis of the potential experimental error sources. Two critical accompanying systematical errors, arising from shaker’s movement distortion along the primary vibration direction and gravity coupling of input acceleration caused by the guide rail deflection, are identified and eliminated through a proposed procedure. Experimental tests demonstrate an uncertainty level two orders of magnitude better than the traditional method based on the dividing head.

Highly sensitive biosensor for specific miRNA detection based on cascade signal amplification and magnetic electrochemiluminescence nanoparticles

Herein, magnetic electrochemiluminescence (ECL) nanoparticle Fe3O4@PtPd/Ru(bpy)32+ had been synthesized then been coupled with CRISPR/Cas13a system and Zn2+ dependent DNAzyme to design a novel ECL biosensor for specific detection of microRNA-145 (miRNA). The synthesized multifunctional magnetic nanoluminescent materials Fe3O4@PtPd/Ru(bpy)32+ not only load Ru(bpy)32+ to provide ECL signals, but also can quickly achieve separation and enrichment from complex matrices. In addition, ferrocene (Fc) was used as a quencher in the Ru(bpy)32+/tripropylamine (TPA) system. Fc was modified on DNA bound to Fe3O4@PtPd. Benefited from the highly specific recognition ability of CRISPR/Cas13a, the target miRNA induces CRISPR/Cas13a trans-cleavage to trigger the Zn2+-dependent DNAzyme cyclic cleavage to realize the dual signal amplification. DNA modified by Fc was split by target miRNA-induced cleaving, and then magnetic separation was performed to keep Fc away from the surface of the nanoparticles. Thus, the enhanced ECL signal was obtained to detect miRNA-145. Under optimized conditions, the prepared sensor showed a wide linear range (1 fM to 1 nM) and a low limit of detection (LOD) down to 0.41 fM. Furthermore, it shows excellent selectivity and good reproducibility. The proposed ECL platform has huge potential applications in the development of various sensitive sensors for detecting the other miRNA.

Ultrasensitive detection of genetic variation based on dual signal amplification assisted by isothermal amplification and cobalt oxyhydroxide nanosheets/quantum dots.

PML/RARα fusion gene (P/R) is the characteristic signature genetic variation of acute promyelocytic leukemia (APL). Here, by integrating triple-stranded DNA hybridization-triggered strand displacement amplification (tri-HT SDA) and cobalt oxyhydroxide nanosheets/quantum dots (CoOOH/QD)-based amplification, we constructed a novel biosensor of easy-operating, time-saving and high sensitivity for detecting P/R to meet clinical needs. Owing to the specific recognition and efficient amplification of tri-HT SDA as well as impressive anti-interference and considerable amplification of CoOOH/QD, this biosensor demonstrated a wide dynamic range (10 fM to 10nM) with a low limit of detection (5.50 fM) in P/R detection. Additionally, this biosensor could detectP/R spiked into human serum with good recoveries and relative standard deviation (RSD), thus potentially exhibiting ultrasensitive and specific nuclear acid sequence detection ability in clinical diagnosis owing to combing isothermal amplification and nanomaterials.

Dual CAR-T Cells Targeting CD19 and CD37 Are Effective in Target Antigen Loss B-cell Tumor Models.

Chimeric antigen receptor T (CAR-T) cells targeting multiple antigens (Ag), may reduce the risk of immune escape following the loss of the target Ag and further increase the efficacy of treatment. We developed dual-targeting CAR-T cells that target CD19 and CD37 Ags and evaluated their antitumor effects. CD19/CD37 dual CAR-T cells were generated using cotransduction and simultaneous gene transfer of two types of lentiviral vectors transferring CD19CAR or CD37CAR genes, including the intracellular domains of CD28 and CD3ζ signaling domains. These dual CAR-T cells contained three fractions: CD19/CD37 bispecific CAR-T cells, single CD19CAR-T cells, and single CD37CAR-T cells. In the functional evaluation of CAR-T cells in vitro, CD19/CD37 dual CAR-T cells showed adequate proliferation and cytokine production in response to CD19 and CD37 antigen stimulation alone or in combination. Evaluation of intracellular signaling revealed that dual CAR-T cell-mediated signals were comparable with single CAR-T cells in response to CD19- and CD37-positive B-cell tumors. Although the cytotoxicity of CD19/CD37 dual CAR-T cells in both CD19- and CD37-positive B-cell tumors was similar to that of single CD19 and CD37CAR-T cells, against CD19 and CD37 Ag-heterogeneous tumor, dual CAR-T cells demonstrated significantly superior tumor lysis compared with single CAR-T cells. Furthermore, CD19/CD37 dual CAR-T cells effectively suppressed Ag-heterogeneous Raji cells in a xenograft mouse model. Collectively, these results suggest that CD19/CD37 dual CAR-T cells may be effective target-Ag-loss B-cell tumor models in vitro and in vivo, which represents a promising treatment for patients with relapsed/refractory B-cell malignancies.

Endogenous CO imaging in bacterial pneumonia with a NIR fluorescent probe

Bacterial pneumonia is a serious respiratory illness that poses a great threat to human life. Rapid and precise diagnosis of bacterial pneumonia is crucial for symptomatic clinical treatment. Endogenous carbon monoxide (CO) is regarded as a significant indicator of bacterial pneumonia; herein, we developed a near-infrared (NIR) probe for fluorescence and photoacoustic (PA) dual-mode imaging of endogenous CO in bacterial pneumonia. NO2-BODIPY could rapidly and specifically react with CO to produce strong NIR fluorescence as well as ratiometric PA signals. NO2-BODIPY has outstanding features including fast response, fluorescence/PA dual mode signals, good specificity, and a low limit of detection (LOD = 20.3 nM), which enables it to image endogenous CO in cells and bacterial pneumonia mice with high sensitivity and high contrast ratio. In particular, NO2-BODIPY has two-photon excited (1340 nm, σ1 = 1671 GM) NIR fluorescence and has been utilized to image endogenous CO in bacterial pneumonia mice with deep tissue penetration. NO2-BODIPY has been demonstrated a good capability of fluorescence/PA dual-mode imaging of CO in bacterial pneumonia mice, providing a precise manner to diagnose bacterial pneumonia.

Engineering ATP activated reversible nanochannel for gating of ion transport

Adenosine triphosphate (ATP) is an important purine molecule. It widely exists in cells and is closely related to a variety of significant physiological and pathological processes, such as energy metabolism, respiratory regulation, inflammation, and so forth. In vivo, ATP not only provides energy for organisms at the molecular level but also activates intracellular ion channels and regulates signaling pathways. Therefore, it is of great significance to construct ATP-responsive nanochannels for understanding complex biological processes. In this work, we designed and fabricated multi-amino conjugated rhodamine B (NH2-RhB) functionalized artificial nanochannels, which exhibited high performance for ATP response with current and fluorescence dual signals. Significantly, owing to the non-covalent bonding of NH2-RhB and ATP, the system performed reversible properties for regulating ion transport and behaved as an ionic gate that can flexibly switch between the "on" and "off" state under the condition of ATP and pH. The platform showed high selectivity for ATP and the detection limit can be as low as 48.2 fM. Moreover, finite-element theoretical simulation was employed to reveal the mechanism of ATP-activated nanochannel regulating ion flux. It proved that the surface charge density of the NH2-RhB functionalized nanochannels can be regulated by dealing with ATP and pH. This work shows promising applications of biomimetic nanochannel with gating properties and offers prospects for the development of biosensing, nanodevice, and nanofluidic logic gates.

A novel electrochemical biosensor based on dual signal amplification of CMK-3@AuNPs and ATRP for DR1 detection

Down-regulator of transcription 1 (DR1) is a crucial biomarker closely related to Hashimoto's Thyroiditis (HT). In this study, a highly selective electrochemical biosensor for ultrasensitive detection of DR1 was prepared by combining nanomaterials CMK-3 @AuNPs and atom transfer radical polymerization (ATRP) signal amplification strategy for the first time. Based on the specific recognition of antigen and antibody, the sensor has strong selectivity. CMK-3 @AuNPs can not only increase the conductivity of the electrode, but also provide more active sites for biological molecules, so as to effectively amplify the output signal. The FMMA monomers were polymerized on the electrode surface through ATRP reaction, which made the electrochemical signal increased significantly. Under optimal conditions, the fabricated electrochemical biosensor showed a wide detection range of 5 × 10-4 ng/mL ∼ 5 × 102 ng/mL with a detection limit as low as 0.159 pg/mL. In addition, the electrochemical biosensor has good stability and anti-interference ability. Therefore, the sensor has broad prospects in the detection of biomarkers.

A strong “on–off” electrochemical biosensor for detecting dicer-like molecules in Fusarium oxysporum f. sp. cubense race 4 (Foc4)

Banana Fusarium wilt is a devastating disease that threatens the global banana industry. The Dicer-like gene (DCL) in race 4 (Foc4) is a key pathogenic factor responsible for the disease. Sensitive and specific detection of DCL is critical in preventing the spread of Foc4 disease. Here, we report the development of a novel electrochemical adapted sensor for the detection of DCL using a ssDNA-hDNA coupled system with a dual signal amplification strategy. The sensor relies on the binding of methylene blue (MB) to both ends of a hairpin probe (hDNA), which is connected to ssDNA through hybridization-mediated, and is linked to the electrode surface via Au-S bonds. The coupling of ssDNA and hDNA with two MBs on each hairpin probe leads to dual signal amplification strategy. In the presence of DCL, hDNA carrying two MBs dissociates from ssDNA, leading to signal attenuation. Compared to sensors using ordinary hairpins, our sensor exhibits larger electrical signals and lower background currents, resulting in a wider detection range (0.01–50 nM) and lower detection limit (12 pM) for DCL. We applied this sensor to the analysis of soil, banana leaves, and fruit samples, and found it to be highly specific and stable. Our method provides a new approach for the detection of banana fusarium wilt and could be useful in assisting with its detection.