Sensitive Detection Of Aflatoxin B1 Research Articles

Bidirectional hybridized hairpin DNA fluorescent aptasensor based on Au-Pd NPs and CDs for ratiometric detection of AFB1.

Anovel and simple ratiometric fluorescent aptasensor was developed for the sensitive detection of aflatoxin B1 (AFB1). A hairpin DNA (h-DNA) was independently synthesized as the basic skeleton, and the bidirectional hybridization of h-DNA can increase the load of aptamer and signal probes, thereby realizing signal amplification. The high-efficiency fluorescence resonance energy transfer interaction between gold-palladium nanoparticles (Au-Pd NPs) and the self-synthesized fluorescent probe carbon dots (CDs) was utilized. Moreover, the label-free probe SYBR Green I (SG I) dye was introduced to form a double-signal probe with CDs, and a ratiometric sensor with FCDs/FSG I as a response signal was constructed. The ratio strategy can eliminate the fluctuation of external factors, thus improving the accuracy and reliability of the sensor. The quenching effect of Au-Pd NPs on CDs was 1.4 times that of AuNPs and 3.4 times that of Pd NPs, respectively. In the range 1-100ng/mL, FCDs/FSG I showed a good linear relationship with the logarithm of the concentration of AFB1, and the limit of detection was as low as 0.07ng/mL. The sensor was used to detect AFB1 in spiked peanuts and wine samples, and the recovery was between 91 and 115%, indicating that the sensor has high application potential in real sample analysis.

Development of tri-mode lateral flow immunoassay based on tailored porous gold nanoflower for sensitive detection of aflatoxin B1

Aflatoxin B1 (AFB1) is regarded as the most toxic mycotoxin and a proven human carcinogen. The development of detection assays for AFB1 is important for human health and environmental protection. In this study, the hollow porous gold nanoflower (HPGN) was synthesized to achieve colorimetric and photothermal performance. Especially, the HPGN exhibited excellent surface-enhanced Raman scattering (SERS) performance by loading with 4-mercaptobenzoic acid (4-MBA), thus SERS determination of AFB1 could be achieved. Compared with gold nanoparticles (AuNPs), the HPGN possessed unique morphology and excellent properties, providing the basis for sensitive and selective AFB1 detection. Under the optimized experimental conditions, the limits of detection (LODs) of lateral flow immunoassay (LFIA) for AFB1 through colorimetric, photothermal and SERS modes were 0.09 ng/mL, 0.09 ng/mL and 0.05 ng/mL, respectively. The tri-mode detection strategy provided a novel concept for immunosensing of mycotoxins or other small molecules.

Colorimetric and ECL dual-mode aptasensor for smartphone-based onsite sensitive detection of aflatoxin B1 in combination with ZnO@MWCNTs/g-C3N4 nanosheets and CuO@CuPt nanocomposites

The development of dual-mode strategies with superior sensitivity and accuracy have garnered increasing attention for researchers in Aflatoxin B1 (AFB1) analysis. Herein, a colorimetric-electrochemiluminescence (ECL) dual-mode biosensor was constructed for onsite and ultrasensitive determination of AFB1. The multi-wall carbon nanotubes (MWCNTs) were integrated with the ZnO metal organic frameworks (MOFs) to accelerate the electron transfer and boost the ECL intensity of g-C3N4 nanoemitters. Through the aptamer-based DNA sandwich assay, the CuO@CuPt nanocomposites were introduced onto the electrode and acted as the dual functional signal nanoprobes. Due to the good spectrum overlap between the CuO@CuPt nanoprobes and g-C3N4 nanosheets, ECL signal could be efficiently quenched. Additionally, the CuO@CuPt nanoprobes show superior catalytic properties towards the TMB and H2O2 colorimetric reactions, and an obvious color alteration from colorless to blue can be observed using the smartphone. Under optimized conditions, a sensitive and accurate dual-mode analysis of the AFB1 was accomplished with the colorimetric detection limit of 3.26 fg/mL and ECL detection limit of 0.971 fg/mL (S/N = 3). This study combines innovative nanomaterial properties of ZnO@MWCNTs, g-C3N4 and CuO@CuPt for ultrasensitive dual-mode detection, which offers new opportunities for the innovative engineering of the dual-mode sensors and demonstrates significant potential in food safety analysis.

In silico studies and development of a protein-based electrochemical sensor for selective and sensitive detection of aflatoxin B1.

Proteins from different species have been dockedwith aflatoxin B1 (AFB1) and identified 3 proteins (prostaglandin-E(2)9-reductase from Oryctolagus uniculus, proto-oncogene serine/threonine-protein kinase Pim-1 and human immunoglobulin G (hIgG)) as potential candidates to develop an electrochemical sensor. Fluorescence spectroscopy experiments have confirmed the interaction of hIgG with AFB1 with an affinity constant of 4.6 × 105 M-1. As a proof-of-concept, hIgG was immobilized on carbon nanocomposite (carbon nanotube-nanofiber, CNT-F)-coated glassy carbon electrode (GCE). FT-IR spectra, HR-TEM and BCA assay have confirmed successful immobilization of hIgG on the electrode (hIgG@CNT-F/GCE). The preparation of this protein electrochemical sensor requires only 1h 36min, which is fast as compared withpreparing an electro immunosensor. hIgG@CNT-F/GCE has displayed an excellent AFB1 limit of detection (0.1 ng/mL), commendable selectivity in the presence of two other mycotoxins (ochratoxin A and patulin) andthe detection of AFB1 in spikedpeanuts and corn samples.

An electrochemical aptasensor based on ACEK enrichment for detection of AFB1

Aflatoxin B1 (AFB1), known for its high toxicity, poses a serious threat to the agriculture and food chain, thereby impacting human health. Herein, a facile electrochemical aptasensor was developed based on alternating current electrokinetic enrichment for rapid and sensitive detection of AFB1. Alternating electric fields on the interdigital electrode array was introduced to improve the sensitivity and time, because the target could be manipulated for migrating from the solution to the sensing interface instead of aimless random motion, leading to the enrichment of the target. The biosensor achieved the trace detection of AFB1, demonstrating a remarkable detection limit of 9.55 × 10−6 ng/mL, and the linear range of 10−3–102 ng/mL. Furthermore, the biosensor was successfully employed for the detection of AFB1 in actual peanuts and peanut oil, demonstrating its great application potential in food analysis.

An ultrasensitive dual-mode approach for AFB1 detection: Colorimetric/label-free SERS aptasensor based on a self-assembled core-shell structured Ag@Au IP6 bifunctional nanozyme

A sensitive and accurate assay of aflatoxin B1 (AFB1) is critically important to ensure food safety. This work designed a dual-mode aptasensor for the convenient and sensitive detection of AFB1 in foodstuff. The aptasensor based on a bifunctional nanozymes was prepared by combining the hybridization chain reaction (HCR) amplification and alkaline phosphatase (ALP)-induced self-assembly reaction. The Ag@Au IP6 nanozyme demonstrated better peroxidase-like activity, effectively initiating the TMB−H2O2 color reaction and facilitating the generation of the blue oxidation product oxTMB. Furthermore, Ag@Au IP6 as a surface-enhanced Raman scattering (SERS) substrate featuring a core-shell structure can notably enhance the Raman signal of oxTMB. When AFB1 existed, the trigger probe exposed and initiated HCR, leading to the introduction of ALP, consequently mediating the generation of Ag@Au IP6 nanozymes, enhancing both SERS signal and colorimetric signal in the TMB-H2O2 system. Consequently, the dual-mode aptasensor is capable of achieving SERS/colorimetric detection with no need for the modification of signal molecules and the replacement of material or reagent. In on-site detection, the visual sensing assay could achieve with collecting and transforming the colorimetric signal to RGB value by a color-assist App in smartphones. Within optimal conditions, the detection limit of dual-mode aptasensor as low as 0.58 pg/L, and with a linear range from 2 pg/L to 200 pg/L, which can meet the needs for AFB1 precise quantification and on-site detection.

In Situ Heterojunction-based PEC Sensor Coupled with LSPR for Sensitive Detection of Aflatoxin B1

To enhance the performance of photoelectrochemical (PEC) sensors, heterojunction construction is a workable technique. However, the interface defects inside the structure are often ignored. In this work, an innovative photoelectrochemical (PEC) sensor based on a “turn-off” PEC model was constructed using the synergistic effects of in situ growing heterojunction and local surface plasmon resonance (LSPR) to detect Aflatoxin B1 (AFB1) instantly. In situ heterojunctions were constructed based on bismuth-based perovskite, and the internal electric field spontaneously established during the growth process can effectively compensate for the interfacial defects and promote the separation and transfer of e−/h+ pairs. Moreover, the gold nanoparticles (Au NPs) LSPR effect can improve PEC performance by enhancing the light trapping ability of the material. The PEC sensor’s detecting area for AFB1 analysis is 10.0 fg/ml-100 ng ml−1, and the detection limit is 4.34 fg ml−1. Good stability, reproducibility, and selectivity underscore the sensor’s suitability for meeting the requirements of food quality monitoring.

Preparation of 3D pomegranate‐like quantum dot nanobeads and its application to highly sensitive detection of aflatoxin B1

Aflatoxin B1 (AFB1) is often used as a major indicator for evaluating aflatoxin contamination in food, and its accurate and sensitive detection is of great significance. Herein, the high-quality CdSe/ZnS fluorescent quantum dot nanobeads (QBs) with 3D pomegranate were prepared by the dual-protecting microemulsion method, which showed excellent stability in different environments. AFB1 were detected by competitive quantum dot-based fluorescence-linked immunosorbent assay (QLISA) using 3D pomegranate-like QBs and single-core silica-encapsulated quantum dots (QD@SiO2). After optimization of the reaction conditions, the quantitative detection range of AFB1 by single-core QD@SiO2 as fluorescent probe was 0–100 ng/mL, with the half maximal inhibitory concentration (IC50) of 3.01 ng/mL and the limit of detection (LOD) of 0.12 ng/mL; whereas the quantitative detection range of AFB1 by fluorescent QBs was 0–100 ng/mL, with the IC50 of 1.76 ng/mL and the LOD of 1.03 pg/mL. Compared to single-core QD@SiO2, the fluorescent QBs was 120-fold more sensitive. The detection of AFB1 was also achieved in negative corn and peanut samples, indicating the possibility of its application to the detection of real food samples. The high performance of competitive QLISA using 3D pomegranate-like QBs as fluorescent probes offer highly sensitive and accurate quantitative detection of analytes in food safety monitoring.

A magnetic graphene oxide and UiO-66 based homogeneous dual recognition electrochemical aptasensor for accurate and sensitive detection of aflatoxin B1

Aflatoxin (AFs) contamination is one of the serious food safety issues. Aflatoxin B1 (AFB1) is the most common and toxic aflatoxin, which has been classified as a class 1 carcinogen by the International Agency for Research on Cancer (IARC). It is extremely destructive to liver tissue. Developing a convenient and sensitive detection technique is essential. In this paper, we developed a homogeneous dual recognition strategy based electrochemical aptasensor for accurate and sensitive detection of aflatoxin B1 (AFB1) based on the magnetic graphene oxide (MGO) and UiO-66. The MGO was synthesized for the recognition and magnetic separation of AFB1 from complex samples. UiO-66/ferrocenecarboxylic acid (Fc)/aptamer composites were constructed as both recognition and signal probes. The probes would specifically capture AFB1 enriched by MGO, which enables dual recognition in homogeneous solution, thus further improving the accuracy of AFB1 detection. The electrochemical aptasensor for AFB1 had a linear range from 0.005 to 500 ng mL−1. Additionally, the limit of detection was 1 pg mL−1. It shows a favorable potential for both sensitive and accurate detection of AFB1 in real samples.

Click Reaction-Mediated Fluorescent Immunosensor Based on Cu-MOF Nanoparticles for Ultrasensitive and High-Throughput Detection of Aflatoxin B1 in Food Samples.

Due to the high toxicity of aflatoxin B1 and its risks to human health, we developed a click reaction-mediated automated fluorescent immunosensor (CAFI) for sensitive detection of aflatoxin B1 based on the Cu(I)-catalyzed click reaction. With its large specific surface area, a copper-based metal-organic framework (Cu-MOF) was synthesized to adsorb and enrich the copper ion (Cu(II)) and then load the complete antigen (BSA-AFB1). After the immunoreaction, Cu(II) inside the Cu-MOF-Antigen conjugate would be reduced to Cu(I) in the presence of sodium ascorbate, which triggered the click reaction between the fluorescent donor-modified DNA and the receptor-modified complementary DNA to lead to a fluorescence signal readout. The whole reaction steps were finished by the self-developed automated immunoreaction device. This CAFI method showed a limit of detection (LOD) of 0.48 pg/mL as well as a 670-fold enhancement in sensitivity compared to conventional ELISA, revealing its great potential in practical applications and automated detection.

Highly sensitive detection of aflatoxin B1 byCRISPR/Cas12a-assisted single nanoparticle counting

CRISPR (clustered regularly interspaced short palindromic repeats) and CRISPR-associated protein (Cas) have gained extensive applications in bioassays. However, CRISPR-based detection platforms are often hampered by limited analytical sensitivity, while nucleic acid-based amplification strategies are usually indispensable for additional signal enhancement with potential risks of amplification leakages. To address these challenges, an amplification-free CRISPR-based bioassay of aflatoxin B1 (AFB1) was proposed by applying single nanoparticle counting. Single-particle mode inductively coupled plasma mass spectrometry (Sp-ICPMS) has been regarded as a sensitive tool for nanoparticle counting since one nanoparticle can generate considerable signals above backgrounds. With AFB1, activator strands were introduced to initiate the trans-cleavage of CRISPR/Cas12a for cutting the nanoparticles-tagged-magnetic beads, which were transduced to nanoparticle count signals after separation. Finally, a pico-mole level limit-of-detections (LODs) with moderate selectivity was achieved. Certified reference materials (CRMs) analysis and recovery tests were conducted with promising results. To our best knowledge, this is the first report of the single particle counting-based CRISPR/Cas12a biosensing study.

A novel electrochemical aptasensor based on AuNPs-loaded graphene oxide-polyaniline for sensitive detection of aflatoxin B1

A novel electrochemical aptasensor based on AuNPs-loaded graphene oxide-polyaniline for sensitive detection of aflatoxin B1

Incorporation of ZnIn2S4 semiconductors with S-vacancy engineered MoS2 nanosheets to develop sensitive photoelectrochemical aptasensor for aflatoxin B1 detection

Aflatoxin B1 (AFB1) has been categorized as group I potent carcinogen to human. Therefore, sensitive detection of AFB1 is of great significance for food safety. Here, a sensitive photoelectrochemical (PEC) aptasensor for AFB1 detection has been developed based on the incorporation of ZnIn2S4 semiconductors with S-vacancy engineered MoS2 (V-MoS2) nanosheets. V-MoS2 nanosheets were obtained by a mild H2O2-mediated chemical etching strategy with varying etching time. ZnIn2S4/V-MoS2 nanocomposites were prepared by a solvothermal method using V-MoS2 as one of the starting materials. Research demonstrated that the incorporation of ZnIn2S4 semiconductors with V-MoS2 could produce ZnIn2S4/V-MoS2 nanocomposites having a broader light absorption range and narrower energy bandgap, which help to generate a larger and highly stable PEC responses. The photocurrent intensity was 3.2 and 5.6 times greater than ZnIn2S4/MoS2 and ZnIn2S4, respectively. Based on the top performing ZnIn2S4/V-MoS2 nanocomposites and the specific aptamer, a PEC aptasensor was successfully constructed for AFB1 detection with a wide linear range of 0.05–50 ng/mL and a low detection limit of 17 pg/mL (S/N = 3). The significance of this work lied not only in representing substantive progress in food safety monitoring, but also in promoting the design of more efficient ZnIn2S4 or V-MoS2-related heterostructures for PEC sensors.

A toluidine blue/porous organic polymer/2D MoSe2 nanocomposite as an electrochemical signaling platform for a sensitive label-free aflatoxin B1 bioassay in some crops

A label-free electrochemical immunosensor using a toluidine blue (TB)/porous organic polymer (POP)/two-dimensional molybdenum diselenide (2D MoSe2) nanocomposite is developed for highly sensitive detection of aflatoxin B1 (AFB1) in selected crops. A POP/2D MoSe2 composite material is employed to modify the surface of a screen-printed carbon electrode (SPCE). Subsequently, TB is adsorbed on the modified SPCE surface, and the resulting TB/POP/2D MoSe2 composite is then used to construct a biosensor. The new POP/2D MoSe2 nanocomposite offers a high surface-to-volume area and is a good electroactive and biocompatible adsorbent for loading TB probe and capture antibodies. Adsorbed TB onto the POP/2D MoSe2 nanocomposite is utilized as a redox probe for the signal amplification unit. This TB/POP/2D MoSe2 nanocomposite provides good electron transfer properties of TB redox probe, good electrical conductivity, good biocompatibility, and likable adsorption ability, thus obtaining a sufficient immobilization quantity of antibodies for the sensor construction. After immobilization of the anti-AFB1 antibody and blocking with BSA on the composite surface, the immunosensor is obtained for the detection of AFB1. Under optimum conditions, the sensor shows a linear logarithmic range of 2.5–40 ng mL−1 with a limit of detection (LOD) of 0.40 ng mL−1. The developed sensor provides several advantages in terms of simplicity, low cost, short analysis time, high selectivity, stability, and reproducibility. Additionally, the proposed immunosensor is successfully validated by the detection of AFB1 in rice, corn, and peanut samples. Utilizing the TB/POP/2D MoSe2 nanocomposite, this label-free electrochemical immunosensor demonstrates outstanding sensitivity and selectivity in detecting AFB1, making it a valuable tool for ensuring the safety of agricultural products and enhancing food security.

UiOL@AIEgens-assisted lateral flow immunosensor for the ultrasensitive dual-modal point-of-care detection of aflatoxin B1

Aflatoxin B1 (AFB1) contamination in food has attracted worldwide attention. The sensitive detection of AFB1 is vital for ensuring food quality and safety. This study developed an ultrasensitive signal-enhanced lateral flow immunosensor (LFIS) based on the functionalized zirconium metal-organic framework (MOF) of a UiO linker enriched with abundant aggregation-induced emission luminogen (UiOL@AIEgens) probes for the rapid dual-modal point-of-care (POC) determination of AFB1. Using UiO MOFs with numerous active sites as the carrier facilitated abundant AIEgens enrichment on the surface. After coupling with enough anti-AFB1 monoclonal antibodies (mAbs), the green-emissive UiOL@AIEgens-mAbs probes with high specificity and remarkably-enhanced fluorescence responses were obtained to competitively capture target AFB1 in the standard or sample solution and AFB1 antigen immobilized on the test (T) line of the POC LFIS. Under optimum conditions, the LFIS was capable of visual qualitative and smartphone-assisted dual-modal determination of target AFB1 within 7 min. Detection occurred in a range of 0.01–5 ng/mL at an ultra-low detection limit of 0.003 ng/mL, which was 300- and 600-fold lower than traditional immunoassays and the maximum limit set by the European Union, respectively. Moreover, the feasibility and robustness of the LFIS platform were assessed by detecting AFB1 in maize and lotus seed samples with average recoveries of 94.3–109.0%. The developed UiOL@AIEgens-based POC LFIS can be used for ultrasensitive, reliable, on-site detection in food. This study provides a new method for the real-time monitoring of AFB1 and other harmful contaminants in food and more complex matrices.

Fe-N-C single-atom nanozyme-linked immunosorbent assay for quantitative detection of aflatoxin B1

Aflatoxin B1 (AFB1) is a prominent food contaminant known for its high toxicity. Rapid, low-cost, and sensitive detection of AFB1 is an urgent demand for food safety risk control. In the traditional enzyme-linked immunosorbent assay (ELISA) method for AFB1 detection, horseradish peroxide (HRP) is high-cost and possesses poor environmental stability. In this study, a direct competitive single-atom nanozymes (SAzymes) -linked immunosorbent assay was developed based on Fe-N-C SAzymes. The proposed methods exhibited excellent sensitivity, rapid detection, fabrication, and low cost. Under the optimal conditions, the limit of detection of the new assay was 3.3 pg mL−1, with a quantitative relationship between 0.0084 and 0.358 ng mL−1. In addition, the developed method presented good recoveries in spiked samples, ranging from 84.50% and 103.75%, with coefficients of variation below 10%. Furthermore, the developed detection method showed good consistency (R2 = 0.9852) with the reference method, liquid chromatography/mass spectrometry (LC-MS/MS). To our knowledge, this is the first report about single-atom nanozymes-based ELISA for AFB1 determination. This work provides a feasible strategy for the rapid and sensitive detection of food contaminants using SAzymes.

A dual-modal biosensor coupling cooperative catalysis strategy for sensitive detection of AFB1 in agri-products.

Herein, the cooperative catalysis effect between nanocomposite (AgPd NPs/POD-M/PEI-rGO) and horseradish peroxidase (HRP) was applied for the fast and sensitive detection of aflatoxin B1 (AFB1). Upon specific and competitive binding of HRP@DNA and AFB1 to cDNA, the working electrode presented different catalytic capacities for supporting electrolytes (TMB and H2O2). In the redox mechanism of TMB and H2O2, HRP and nanocomposite effectively catalyzed the oxidization of TMB to form the one-electron oxidation intermediate TMB+, and contributed the electrical signals and absorbance signals. Electrochemistry and colorimetric analyses were successfully realized for AFB1 detection with 0.2pg/mL and 8pg/mL of detection limits, respectively, which is much lower than that of traditional HPLC methods. Overall, this method had significant reliability and sensitivity, offering a promising potential for conveniently evaluating the quality of agri-products polluted with AFB1. Moreover, this approach provides a new idea for fast and accurate detection of mycotoxin.

Self-Assembly Multivalent Fluorescence-Nanobody Coupled Multifunctional Nanomaterial with Colorimetric Fluorescence and Photothermal to Enhance Immunochromatographic Assay.

The multimodal lateral flow immunoassay (LFIA) has provided accurate and reliable results for fast and immediate detection. Nonetheless, multimodal LFIA remains challenging to develop biosensors with high sensitivity and tolerance to matrix interference in agro-food. In this study, we developed a self-assembled multivalent fluorescence-nanobody (Nb26-EGFP-H6) with 16.5-fold and 30-fold higher affinity and sensitivity than a monovalent nanobody (Nb26). Based on the Nb26-EGFP-H6, we synthesized enhanced immune-probes Zn-CN@Nb26-EGFP-H6 by pyrolyzing and oxidizing an imidazolating zeolite framework-8 (ZIF-8) to obtain photothermal metal-carbon nanomaterials (Zn-CN) for immobilizing Nb26-EGFP-H6. The rough and porous structure of Zn-CN with a large surface area facilitates the enrichment and immobilization of antibodies. A trimodal lateral flow immunoassay (tLFIA) with colorimetric, fluorescent, and photothermal triple signal outputs was constructed for the detection of aflatoxin B1 (AFB1) in maize. Attractively, the Zn-CN-based tLFIA's multiplex guarantees accurate and sensitive detection of AFB1, with triple signal detection limits of 0.0012 ng/mL (colorimetric signals), 0.0094 ng/mL (fluorescent signals), and 0.252 ng/mL (photothermal signals). The sensitivity of the trimode immunosensor was 628-fold and 42-fold higher than that of the original Nb26-based ELISA (IC50) and the unimodal LFIA (LOD). This work provides an idea for constructing a sensitive, tolerant matrix and efficient and accurate analytical platform for rapidly detecting AFB1 in food.

MnO2 nanoflowers based colorimetric and fluorescent dual-mode aptasensor for sensitive detection of aflatoxin B1 in milk

Aflatoxin B1 (AFB1) with tremendous toxic effects has caused a serious threat to food security. Accurate quantification of AFB1 in food can effectively prevent the risk of human intake of AFB1. Herein, a colorimetric and fluorescent dual-mode aptasensor for accurate and sensitive detection of AFB1 has been developed based on MnO2 nanoflowers (MnO2NFs) for the first time. MnO2NFs could catalyze the oxidation of 3,3′,5,5′-tetramethylbenzidine (TMB) into blue oxidation product (TMBox) by H2O2, which would be used for visible detection of AFB1. Simultaneously, MnO2NFs can be served as a signal amplifier and reduced by ascorbic acid to generate lots of Mn2+ which would quench the fluorescence of calcein for fluorescent detection of AFB1. Both colorimetric and fluorescent methods have been successfully applied for determination of AFB1 in milk samples with satisfactory results. The proposed dual-mode detection method with high detection sensitivity and accuracy showed great promise for monitoring AFB1 in food.

Accelerated Hybridization Chain Reaction Kinetics Using Poly DNA Tetrahedrons and Its Application in Detection of Aflatoxin B1.

Traditional hybridization chain reaction (HCR) as a popular isothermal amplification technique shows some inevitable disadvantages in bioanalysis due to its relatively slow kinetics, which could be markedly promoted when the HCR initiator occurs under tension. Herein, a poly DNA tetrahedrons (pTDNs)-mediated HCR was successfully constructed to make its initiator in a stretched state by long-range electrostatic forces owing to the superimposed electrostatic interactions derived from the synthesized pTDNs, and it was hypothesized that it could remarkably enhance HCR performance, which was testified by theoretical simulations and experimental studies. Consequently, pTDNs-mediated HCR was applied to develop a novel immunoassay for rapid and sensitive detection of aflatoxin B1 as a proof-of-concept, and its signal amplification was attributed to the increased G4 DNAzyme that loaded on the second antibody. Our work paves a promising way using simple DNA frameworks alone to heighten HCR kinetics for reaction speed improvement and signal amplification in bioanalysis.