Electrochemical Aptasensor Research Articles

Surface‐Based Multimeric Aptamer Generation and Bio‐Functionalization for Electrochemical Biosensing Applications

AbstractMultimeric aptamers have gained more attention than their monomeric counterparts due to providing more binding sites for target analytes, leading to increased affinity. This work attempted to engineer the surface‐based generation of multimeric aptamers by employing the room temperature rolling circle amplification (RCA) technique and chemically modified primers for developing a highly sensitive and selective electrochemical aptasensor. The multimeric aptamers, generated through surface RCA, are hybridized to modified spacer primers, facilitating the positioning of the aptamers in the proximity of sensing surfaces. These multimeric aptamers can be used as bio‐receptors for capturing specific targets. The surface amplification process was fully characterized, and the optimal amplification time for biosensing purposes was determined, using SARS‐CoV‐2 spike protein (SP). Interestingly, multimeric aptasensors produced considerably higher response signals and affinity (more than 10‐fold), as well as higher sensitivity (almost 4‐fold) compared to monomeric aptasensors. Furthermore, the impact of surface structures on the response signals was studied by utilizing both flat working electrodes (WEs) and nano‐/microislands (NMIs) WEs. The NMIs multimeric aptasensors showed significantly higher sensitivity in buffer and saliva media with the limit of detection less than 2 fg/ml. Finally, the developed NMIs multimeric aptasensors were clinically challenged with several saliva patient samples.

Surface-Based Multimeric Aptamer Generation and Bio-Functionalization for Electrochemical Biosensing Applications.

Multimeric aptamers have gained more attention than their monomeric counterparts due to providing more binding sites for target analytes, leading to increased affinity. This work attempted to engineer the surface-based generation of multimeric aptamers by employing the room temperature rolling circle amplification (RCA) technique and chemically modified primers for developing a highly sensitive and selective electrochemical aptasensor. The multimeric aptamers, generated through surface RCA, are hybridized to modified spacer primers, facilitating the positioning of the aptamers in the proximity of sensing surfaces. These multimeric aptamers can be used as bio-receptors for capturing specific targets. The surface amplification process was fully characterized, and the optimal amplification time for biosensing purposes was determined, using SARS-CoV-2 spike protein (SP). Interestingly, multimeric aptasensors produced considerably higher response signals and affinity (more than 10-fold), as well as higher sensitivity (almost 4-fold) compared to monomeric aptasensors. Furthermore, the impact of surface structures on the response signals was studied by utilizing both flat working electrodes (WEs) and nano-/microislands (NMIs) WEs. The NMIs multimeric aptasensors showed significantly higher sensitivity in buffer and saliva media with the limit of detection less than 2 fg/ml. Finally, the developed NMIs multimeric aptasensors were clinically challenged with several saliva patient samples.

Highly conductive AuNPs/Co-MOF nanocomposites synergistic hybridization chain reaction enzyme-free electrochemical aptasensor for ultrasensitive detection of Aflatoxin B1

Aflatoxin B1 (AFB1) is the most poisonous and carcinogenic pollutant among numerous mycotoxins, posing a serious risk to the health of humans. Herein, an electrochemical aptasensor was constructed for AFB1 detection using gold nanoparticles/cobalt-based metal–organic skeleton (AuNPs/Co-MOF) as a modifying material combined with an enzyme-free signal amplification strategy. AuNPs/Co-MOF have an extensive specific surface area and outstanding electrical conductivity, which allows for the loading of a large number of DNA strands and enhancement of the thionine (THi) redox signal, leading to a significant increase in signal response THi loaded bimetallic core–shell nanocomposites (Au@PtNPs) were synthesized as signal labels. Au@PtNPs have high catalytic activity towards THi while improving the efficiency of the loaded THi, which enables the electrochemical signal of THi to be amplified. In addition, the hybridization chain reaction (HCR) signal amplification strategies for enzyme-free amplification elicits a strong response in the electrochemical signal while ensuring sensor stability. The aptasensor has a detection range of 0.001–500 ng/mL and achieves the limit of detection (LOD) was 1.2 × 10-2 pg/mL and limit of quantification (LOQ) was 4.0 × 10-2 pg/mL under optimal experimental conditions. Therefore, applications for electrochemical aptasensors in the realm of food safety monitoring are quite promising.

Construction of an aptasensor based on the composite nanomaterial IL-rGO-TEPA/SiO2-AuPt and its high-sensitive detection of cardiac troponin I

To accurately diagnose and evaluate the degree of myocardial injury and predict myocardial infarction disease, the rapid and sensitive detection of cardiac troponin I (cTnI) is required. Therefore, a label-less electrochemical aptasensor was prepared for cTnI determination. Ionic liquid 1-hydroxyethy l-3- methyl imidazole four fluorine boric acid salt (IL)-reduced graphene oxide − four ethylene oxidation reduction five amine (rGO-TEPA)/silica (SiO2)- gold platinum (AuPt) (IL-rGO-TEPA/SiO2-AuPt) nanocomposites were synthetized as the substrate nanocomposite and were characterized using a series of characterization methods. The performance of the prepared aptasensor was characterized through electrochemical methods. Result displayed that better substrate materials were synthesized, and the prepared electrochemical aptasensor showed excellent electrochemical performance. Under optimal conditions, the electrochemical aptasensor had a salient analytical performance. The electrochemical signals linearly decreased with the logarithm of the cTnI concentration between the 0.5 pg·mL−1 and 1 × 106 pg·mL−1 concentration range (R2 value = 0.9955) and the aptasensor had a wide detection range (0.5 ∼ 1 × 106 pg·mL−1) and a low detection limit of 0.4 pg·mL−1. At the same time, the aptasensor had good selectivity, repeatability and stability. Relative standard deviation (RSD) values ranged from 3.45 % ∼ 4.97 % and the recovery rate was 97.5 % ∼ 103 % in human serum samples, the RSD values of the spiked recovery and ELISA method were less than 5 %. This study provides a new theoretical basis and method for the clinical detection of cTnI.

Aptamer-enabled electrochemical bioplatform utilizing surface-modified g-C3N4/MoS2/PANI nanocomposite for detection of CA125 biomarker

Detecting ovarian cancer at an early stage is crucial for enhancing patient outcomes, underscoring the demand for an efficient and non-invasive detection method. Cancer antigen 125 (CA125) is a vital biomarker in ovarian cancer detection, and there is a pressing demand to develop a quick, sensitive, and simple detection method. Nanobiosensors are increasingly being used by scientists due to their high selectivity and sensitivity, allowing for the swift and precise detection of a wide range of biomarkers. This study aimed to design and fabricate an electrochemical nanobiosensor that could accurately and selectively detect CA125. The nanobiosensor employed graphitic carbon nitrides, molybdenum disulfide, and polyaniline (g-C3N4/MoS2/PANI) to stabilize aptamer strands on a modified glassy carbon electrode. The aptasensor was used to perform electrochemical detection of labeled CA125, utilizing methylene blue and label-free ferrocyanide methods. Ferrocyanide and methylene blue detection limits were determined to be 0.196 U.mL−1 for ferrocyanide and 0.196 U.mL−1 for methylene blue, with a linear detection range of 2–10 U.mL−1 for linear detection. The study results showed that the modified electrode exhibited high selectivity towards CA125 and superior stability compared to other biomolecules. The electrochemical aptasensor also displayed impressive performance when analyzing the serum of patients and healthy people. These findings hold significant promise for future investigation in ovarian cancer diagnosis. This novel electrochemical nanobiosensor may aid in the early detection and management of ovarian cancer, ultimately leading to better patient outcomes.

A microfluidic ratiometric electrochemical aptasensor for highly sensitive and selective detection of 3,3',4,4'-tetrachlorobiphenyl.

This study proposes a strategy using a microfluidic ratiometric electrochemical aptasensor to detect PCB77 with excellent sensitivity and specificity. This sensing platform combines a microfluidic chip, a wireless integrated circuit system for aptamer-based electrochemical detection, and a mobile phone control terminal for parameter configuration, identification, observation, and wireless data transfer. The sensing method utilizes a cDNA (MB-COOH-cDNA-SH) that is labelled with the redox probe Methylene Blue (MB) at the 5' end and has a thiol group at the 3' end. Additionally, it utilizes a single strand PCB aptamer that has been modified with ferrocenes at the 3' end (aptamer-Fc). Through gold-thiol binding, the labelled probe of MB-COOH-cDNA-SH was self-assembled onto the surface of an Au/Nb2CTx/GO modified electrode. On exposure to aptamer-Fc, it will hybridize with MB-COOH-cDNA-SH to form a stable double-stranded structure on the electrode surface. When PCB77 is present, aptamer-Fc binds specifically to the target, enabling the double-stranded DNA to unwind. Such variation caused changes in the differential pulse voltammetry (DPV) peak currents of both MB and Fc. A substantial improvement is observed in the ratio between the two DPV peaks. Under the optimum experimental conditions, this assay has a response that covers the 0.0001 to 1000 ng mL-1 PCB77 concentration range, and the detection limit is 1.56 × 10-5 ng mL-1. The integration of a ratiometric electrochemical aptasensor with designed microfluidic and integrated devices in this work is an innovative and promising approach that offers an efficient platform for on-site applications.

Construction of pitch-derived porous carbon-functionalized electrochemical aptasensor for sensitivity detection of chloramphenicol

Construction of pitch-derived porous carbon-functionalized electrochemical aptasensor for sensitivity detection of chloramphenicol

Chronocoulometric Quantum Dot Aptasensor for SARS‐CoV‐2 Nucleocapsid Protein Biomarker

AbstractFor the first time, a novel mercaptosuccinic acid‐capped tungsten telluride quantum dot (MSA‐WTe3‐QD)‐based electrochemical aptasensor was designed for the determination of SARS‐CoV‐2 N‐protein in synthetic human serum, plasma and urine. The MSA‐WTe3‐QD which exhibited spherical morphology and an average diameter of 3.56 nm were synthesized through a bottom‐up approach. The aptasensor was prepared by firstly immobilizing MSA‐WTe3‐QD on a screen‐printed carbon electrode (SPCE) by using the 1‐ethyl‐3‐(3‐dimethylaminopropyl)carbodiimide/N‐hydroxysuccinimide (EDC/NHS) coupling chemistry. Thereafter, amine‐terminated 90‐mer ssDNA aptamer was deposited on the modified SPCE and left to cure to form the aptasensor. Using the chronocoulometry signal transduction technique, the aptasensor's calibration with N‐protein gave a linear range value of 1–9 pg.mL−1 and a limit of detection (LOD) value of 0.08 pg.mL−1. The LOD value of the aptasensor was 8 times lower than the LOD value of 0.71 pg.mL−1 obtained with a commercial N‐protein ELISA kit. When tested in simulated real samples the aptasensor recovery rates of 95–103.33 % were obtained. In the presence of various interferents, the aptasensor displayed good selectivity to N‐protein.

Simultaneous detection of multiple mycotoxins using MXene-based electrochemical aptasensor array and a self-developed multi-channel portable device

In response to the pressing need for highly efficient simultaneous detection of multiple mycotoxins, which are often found co-occurring in food raw materials and feed, an MXene-based electrochemical aptasensor array (MBEAA) was developed. This aptasensor array utilizes high-specificity aptamers as recognition elements, enabling the capture of electrical signal changes in the presence of target mycotoxins. Based on this platform, a multi-channel portable electrochemical device, enabling rapid, cost-effective, and simultaneous detection of aflatoxin B1 (AFB1), ochratoxin A (OTA), and zealenone (ZEN) was further developed. The developed system boasts a wide detection range of 1.0 × 10−1 to 10.0 ng mL−1, with remarkable performance characterized by ultra-low detection limits of 41.2 pg mL−1, 27.6 pg mL−1, and 33.0 pg mL−1 for AFB1, OTA, and ZEN, respectively. Successfully applied in corn samples, this method offers a portable, easy-to-operate, and cost-effective solution for simultaneous multi-mycotoxin detection. Moreover, the application of the self-developed detection system could be expanded for simultaneous detection of many different targets when their specific aptamers or antibodies were available.

Label-free electrochemical aptasensor for the detection of HepG2 hepatocellular carcinoma cells

Hepatocellular carcinoma (HCC) is the most common liver malignancy and is characterized by increasing incidence and high mortality rates. Current methods for the screening and diagnosis of HCC exhibit inherent limitations, highlighting the ever-growing need for the development of new methods for the early diagnosis of HCC. The aim of this work was to develop a novel electrochemical aptasensor for the detection of HepG2 cells, a type of circulating tumor cells that can be used as biomarkers for the early detection of HCC. A carbon screen-printed electrode was functionalized with a composite suspension containing graphene oxide, chitosan, and polyaniline nanoparticles to increase the electrode surface and provide anchoring sites for the HepG2 cell-specific aptamer. The aptamer was immobilized on the surface of the functionalized electrode using multipulse amperometry, an innovative technique that significantly reduces the time required for aptamer immobilization. The innovative platform was successfully employed for the first time for the amplification-free detection of HepG2 cells in a linear range from 10 to 200,000 cells/mL, with a limit of detection of 10 cells/mL. The platform demonstrated high selectivity and stability and was successfully used for the detection of HepG2 cells in spiked human serum samples with excellent recoveries.Graphical abstract

Cell-SELEX for aptamer discovery and its utilization in constructing electrochemical biosensor for rapid and highly sensitive detection of Legionella pneumophila serogroup 1

This study introduces an innovative electrochemical aptasensor designed for the highly sensitive and rapid detection of Legionella pneumophila serogroup 1 (L. pneumophila SG1), a particularly virulent strain associated with Legionellosis. Employing a rigorous selection process utilizing cell-based systematic evolution of ligands by exponential enrichment (cell-SELEX), we identified new high-affinity aptamers specifically tailored for L. pneumophila SG1. The selection process encompassed ten rounds of cell-SELEX cycles with live L. pneumophila, including multiple counter-selection steps against the closely related Legionella sub-species. The dissociation constant (Kd) of the highest affinity sequence to L. pneumophila SG1 was measured at 14.2 nM, representing a ten-fold increase in affinity in comparison with the previously reported aptamers. For the development of electrochemical aptasensor, a gold electrode was modified with the selected aptamer through the formation of self-assembled monolayers (SAMs). The newly developed aptasensor exhibited exceptional sensitivity, and specificity in detecting and differentiating various Legionella sp., with a detection limit of 5 colony forming units (CFU)/mL and an insignificant/negligible cross-reactivity with closely related sub-species. Furthermore, the aptasensor effectively detected L. pneumophila SG1 in spiked water samples, demonstrating an appreciable recovery percentage. This study shows the potential of our aptamer-based electrochemical biosensor as a promising approach for detecting L. pneumophila SG1 in diverse environments.

Aptamer and Electrochemical Aptasensor towards Selenate Ions (SeO42-).

Selenium is an essential inorganic compound in human and animal nutrition, involved in the proper functioning of the body. As a micronutrient, it actively contributes to the regulation of various metabolic activities, i.e., thyroid hormone, and protection against oxidative stress. However, Se exhibits a narrow concentration window between having a positive effect and exerting a toxic effect. In higher doses, it negatively affects living organisms and causes DNA damage through the formation of free radicals. Increased reactivity of Se anions can also disrupt the integrity and function of DNA-repairing proteins. As the permissible concentration of Se in drinking water is 10 µg/L, it is vital to develop sensitive and robust methods of Se detection in aqueous samples. In this study, for the first time, we proposed a selective aptamer for selenate ion detection, chosen following the SELEX process, and its application in the construction of an electrochemical aptasensor towards SeO42- ions. Measurement conditions such as the used redox marker and pH value of the measurement solution were chosen. The proposed aptasensor is characterized by good selectivity and an LOD of 1 nM. Conditions for biosensor regeneration and storage were also investigated in this research.

Competitive electrochemical aptasensor for high sensitivity detection of liver cancer marker GP73 based on rGO-Fc-PANi nanocomposites

Golgi protein 73 (GP73) is a novel tumor marker in the early diagnosis and prognosis of hepatocellular carcinoma (HCC). Herein, a competitive electrochemical aptasensor for detecting GP73 was constructed using reduced graphene oxide-ferrocene-polyaniline nanocomposite (rGO-Fc-PANi) as the biosensing platform. The rGO-Fc-PANi had larger specific surface area, excellent conductivity and outstanding electroactive performance, which served as nanocarrier for GP73 aptamer (GP73Apt) binding and as redox nanoprobe for record electrical signal. Then, a complementary chain (cDNA) was fixed to the electrode by hybridization with GP73Apt. When GP73 was present, a competitive process happened among cDNA, GP73Apt and GP73, formed the GP73-GP73Apt stable chemical structure and made cDNA detach from the sensing electrode, resulting in enhancement of electrical signal. The difference in the corresponding peak current before and after the competition can be used to indicate the quantitative of GP73. Under optimal conditions, the DPV current response showed a good log-linear relationship with GP73 concentrations (0.001 ∼ 100.0 ng/mL) with a detection limit of 0.15 pg/mL (S/N = 3). It was successfully used for GP73 detection in human serum with RSDs ranging from 1.08 % to 6.96 %. Therefore, the aptasensor could provide an innovative technology platform and hold a great potential in clinical application.

Designing an electrochemical aptasensor based on a microstructure porous-covalent organic polymer for the determination of 2,4,6-trinitrotoluen in water and soil samples

A selective electrochemical aptasensor assessment based on a microstructure porous-covalent organic polymer (MP-COP) was introduced and applied for 2,4,6-Trinitrotoluen (TNT) sensing. The synthesis of MP-COP involves the formation of an amide bond by coupling carboxylic acids and amine groups using trimesic acid. The synthesized MP-COP was used to modify the glassy carbon electrode (GCE) due to its ability to enhance electrochemical activity. The GCE is modified with the MP-COP and aptamer to create a sensing substrate, with the goal of improving the aptasensor’s electrochemical performance for TNT detection. Electrochemical impedance spectroscopy, differential pulse voltammetry, and cyclic voltammetry were applied for the evaluation of the introduced aptasensor in [Fe(CN)6]3−/4− probe. Moreover, it was delineated using fourier-transform infrared spectroscopy, X-ray diffraction, scanning electron microscopy, Brunauer-Emmett-Teller. The suggested aptasensor for TNT has a limit of detection of 0.0003 pM and the linear range from 0.001 to 100 pM. The usability of the proposed aptasensor for TNT measurement in actual samples was examined with the recovery range from 98.30 to 102.40 %. It seems that the proposed strategy can be expanded to design of electrochemical aptasensors.

Ratiometric electrochemical aptasensor with strand displacement for insulin detection in blood samples

BackgroundDiabetes patients suffer either from insulin deficiency or resistance with a high risk of severe long-term complications, thus the quantitative assessment of insulin level is highly desired for diabetes surveillance and management. Utilizing insulin-capturing aptamers may facilitate the development of affordable biosensors however, their rigid G-quadruplex structures impair conformational changes of the aptamers and diminish the sensor signals. ResultsHere we report on a ratiometric, electrochemical insulin aptasensor which is achieved by hybridization of an insulin-capturing aptamer and a partially complementary ssDNA to break the rigid G-quadruplex structures. To improve the durability of the aptasensor, the capturing aptamer was immobilized on gold electrodes via two dithiol-phosphoramidite functional groups while methoxy-polyethylene glycol thiol was used as a blocking molecule. The exposure of the sensor to insulin-containing solutions induced the dissociation of the hybridized DNA accompanied by a conformational rearrangement of the capturing aptamer back into a G-quadruplex structure. The reliability of sensor readout was improved by the adoption of an AND logic gate utilizing anthraquinone and methylene blue redox probes associated to the aptamer and complementary strand, respectively. Our aptasensor possessed an improved detection limit of 0.15 nM in comparison to aptasensors without strand displacement. SignificanceThe sensor was adapted for detection in real blood and is ready for future PoC diagnostics. The capability of monitoring the insulin level in an affordably manner can improve the treatment for an increasing number of patients in developed and developing nations. The utilization of low-cost and versatile aptamer receptors together with the engineering of ratiometric electrochemical signal recording has the potential to considerably advance the current insulin detection technology toward multi-analyte diabetes sensors.

A molecularly imprinted electrochemical aptasensor-based dual recognition elements for selective detection of dexamethasone

In this work, a novel molecularly imprinted electrochemical aptasensor (MIEAS) was developed for highly selective detection of dexamethasone (Dex) in natural water environment. Gold nanoparticles (AuNPs) modified by nitrogen doped molybdenum carbide-graphene (N–Mo2C-Gr) were employed as the supports, where N–Mo2C-Gr improved the conductivity of the electrode and provided a larger specific surface area to polymerize more active substances. Using Dex as template molecule, o-phenylenediamine (o-PD) as the chemical functional monomer and aptamer as the biofunctional monomer, a molecularly imprinted polymer (MIP) membrane with Dex specific recognition sites was formed by electropolymerization. Due to the synergistic effect of MIP and aptamers, the as-prepared MIEAS exhibited a decent linear relationship to Dex detection within a relatively wide range of 10−13 – 10−5 M, and the detection limit was 1.79 × 10−14 M. The recovery in actual water and tablet samples is satisfactory, which confirms the potential application prospects of this sensor in the determination of Dex.

An ultrasensitive electrochemical aptasensor based on zeolitic imidazolate framework-67 loading gold nanoparticles and horseradish peroxidase for detection of aflatoxin B1

Aflatoxin B1 (AFB1) is one of the most toxic mycotoxins and poses a high risk to human health. Highly sensitive and rapid detection is one of the most effective preventive measures to avoid potential hazards. Herein, an electrochemical aptasensor based on DNA nanotetrahedron and zeolitic imidazolate framework-67 loading gold nanoparticles, horseradish peroxidase, and aptamers was designed for the ultrasensitive detection of AFB1. The high specific surface area and large pore volume of zeolitic imidazolate framework-67 can increase the loading capacity and further improve the detection sensitivity of electrochemical aptasensors. DNA nanotetrahedron can enhance the capture ability of AFB1 with steady immobilization. The developed aptasensor showed good analytical performance for AFB1 detection, with a detection limit of 3.9 pg mL−1 and a wide linear range of 0.01–100 ng mL−1. The aptasensor detected AFB1 in corn samples with recovery rates ranging from 94.19%–105.77% and has potential for use in food safety monitoring.

Simultaneous Hg2+ and Pb2+ detection in water samples using an electrochemical aptasensor with dual signal amplification by exonuclease III and metal-organic frameworks

Heavy metal pollution in the environment has become a significant global concern due to its detrimental effects on human health and the environment. In this study, we report an electrochemical aptasensor for the simultaneous detection of Hg2+ and Pb2+. Gold nanoflower/polyethyleneimine-reduced graphene oxide (AuNFs/PEI-rGO) was introduced on the surface of a gold electrode to improve sensing performance. The aptasensor is based on the formation of a T-Hg2+-T mismatch structure and specific cleavage of the Pb2+-dependent DNAzyme, resulting in a dual signal generated by the Exo III specific digestion of methylene blue (MB) labeled at the 3′ end of probe DNA-1 and the reduction of the substrate ascorbic acid (AA) catalyzed by the signal label. The decrease of MB signal and the increase of AA oxidation peak was used to indicate the content of Hg2+ and Pb2+, respectively, with detection limits of 0.11 pM (Hg2+) and 0.093 pM (Pb2+). The aptasensor was also used for detecting Hg2+ and Pb2+ in water samples with good recoveries. Overall, this electrochemical aptasensor shows promising potential for sensitive and selective detection of heavy metals in environmental samples.

A rapid and ultrasensitive cardiac troponin I aptasensor based on an ion-sensitive field-effect transistor with extended gate

Acute myocardial infarction (AMI) is a life-threatening disease with a short course and a high mortality rate. However, it is still a great challenge to achieve the on-site diagnosis of this disease within minutes, meaning there is an urgent need to develop an efficient technology for realizing the rapid diagnosis and early warning of AMI in clinical emergencies. In this study, an ultrasensitive electrochemical aptasensor based on an extended-gate ion-sensitive field-effect transistor (EGISFET) was designed to achieve the quantitative assay of cardiac troponin I (cTnI), which is a highly sensitive and specific biomarker of AMI, within only 5 min. The EGISFET exhibits extremely high detection sensitivity due to its separated structure with a large sensing area and the surface-modified Prussian blue-gold nanoparticles (PB-AuNPs) composite, which serves as a signal magnifier and DNA loading platform for good electrocatalytic ability with a large specific area. Additionally, a target-induced strand-release strategy is proposed to shorten the recognition time of cTnI using a particular DNA strand. Under optimal conditions, the as-prepared aptasensor exhibits a wide linear range of 1–1000 pg/mL, an ultralow detection limit of 0.3 pg/mL, and reliable detection results in real serum samples. It is highly anticipated that this EGISFET-based aptasensor will have broad applications in the early warning and rapid diagnosis of AMI and other acute diseases in emergency treatment.

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.