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# Detection of aflatoxin B1 using DNA sensors: a review

Tianhan Xie, Eric Lichtfouse, Zaina Omary Mochiwa, Jin Wang, Bangxing Han, Li Gao

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## To cite this version:

Tianhan Xie, Eric Lichtfouse, Zaina Omary Mochiwa, Jin Wang, Bangxing Han, et al.. Detection of aflatoxin B1 using DNA sensors: a review. Environmental Chemistry Letters, 2025, 23, pp.1425-1443. 10.1007/s10311-025-01842-7. hal-05038842

HAL Id: hal-05038842

https://hal.science/hal-05038842v1

Submitted on 18 Apr 2025

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# Detection of aflatoxin B1 using DNA sensors: a review

Tianhan Xie<fnref n="1" /> · Eric Lichtfouse<fnref n="2" /> Ⓒ · Zaina Omary Mochiwa<fnref n="1" /> · Jin Wang<fnref n="1" /> · Bangxing Han<fnref n="3" /> · Li Gao<fnref n="1" />,<fnref n="4" /> Ⓒ

## Abstract

Aflatoxin B1 is a mycotoxin produced by the fungus *Aspergillus* that contaminates food, notably grains and peanuts. Aflatoxin B1 is hepatotoxic, causing necrosis, and cirrhosis, and is classified as an hepatocarcinogen. Traditional methods for detecting aflatoxin B1 such as thin-layer chromatography, high-performance liquid chromatography, enzyme-linked immunoassay, and liquid chromatography-tandem mass spectrometry, have limitations including high costs, complex preparation procedures, and occasionally low sensitivity. Here, we review DNA-based biosensors for aflatoxin B1 detection with emphasis on electrochemical and optical sensors. Electrochemical biosensors are based on electrochemical impedance spectroscopy, amperometry, voltammetry, and potentiometry. Optical sensors involve colorimetry, surface plasmon resonance, fluorescence, and electrochemiluminescence. Sensors combine nano and composite materials, such as gold nanoparticles, black phosphorus nanosheets, graphene oxide, niobium carbide, photonic crystals, and liquid crystals. DNA-based biosensors, such as aptamer biosensors, are efficient, rapid, sensitive, affordable, and selective to detecting contaminants and pathogens.

**Keywords** DNA technology · Biosensor · Aflatoxin B1 · Aptamer · Nanomaterial

## Introduction

Aflatoxins are carcinogenic mycotoxins produced by *Aspergillus* species such as *Aspergillus flavus* and *Aspergillus parasiticus* [1]. These toxins were first identified in 1960 as the cause of the “Turkey X disease” in the outskirts of London, England, after it killed hundreds of thousands of turkeys [2]. Aflatoxins are widely present in major food crops, including dried fruits, rice, spices, dried fruits, peanuts, corn, and

dairy products, leading to irreversible food contamination [3–7, 17]. Detecting and screening for aflatoxins is thus essential to prevent food poisoning and safeguard public health [8–10]. There are approximately 20 types of aflatoxins derivatives, named B1, B2, M1, M2, G1, G2, Q1, P1, and GM, with aflatoxin B1 being the most toxic [11, 12].

Aflatoxin B1 is one of the four aflatoxins generated by *Aspergillus* species. Aflatoxin B1 is a difuran cyclic toxin and is currently regarded as the most toxic carcinogen worldwide, with potent carcinogenic and teratogenic effects [13–17]. The primary target of aflatoxin B1 is the liver, where it is metabolized by the microsomal chromoenzyme CYQ450 to form AFB1-8,9-epoxide. This epoxide can then form adducts with amino acids and DNA, potentially causing genetic mutations leading to cancer [18–20]. Aflatoxin B1 is a food contaminant that commonly grows on sorghum, peanuts, and barley. Aflatoxin B1 and its by-products cause irreversible damage to living organisms [21–23].

Aflatoxin B1 has been categorized as a group 1 carcinogenic substance according to the International Agency for Research on Cancer (IARC) of the World Health Organization (WHO) [24]. Numerous countries and regions have established upper thresholds for aflatoxin B1 residues in food, known as maximum residue limits (MRLs). For instance, the limit for corn, peanuts, and related products

<footnote>☒ Eric Lichtfouse
eric.lichtfouse@icloud.com</footnote>

<footnote>☒ Bangxing Han
hanbx1978@sina.com</footnote>

<footnote>☒ Li Gao
gaoli@ujs.edu.cn</footnote>

<footnote n="1">¹ School of Life Sciences, Jiangsu University,
Zhenjiang 212013, China</footnote>

<footnote n="2">² State Key Laboratory of Multiphase Flow in Power
Engineering, Xi’an Jiaotong University, Xi’an 710049,
Shaanxi, People’s Republic of China</footnote>

<footnote n="3">³ College of Biological and Pharmaceutical Engineering, West
Anhui University, Moon Island, Luan 237012, China</footnote>

<footnote n="4">⁴ School of Life Sciences, Qinghai Normal University,
Xi Ning 810000, China</footnote>
in China and the USA is 20 µg/kg [25–27]. The European Commission allows a maximum of 4 µg/kg for AFB1, AFB2, AFG1, and AFG2, and a maximum level of 2 µg/kg aflatoxin B1 in cereals and grain derivatives [28–30].

The timely and reliable aflatoxin B1 identification is essential to ensure consumer safety and to improve the quality of food and crops worldwide. Currently, detection methods mainly include thin-layer chromatography (TLC) [31], high-performance liquid chromatography (HPLC) [32], enzyme-linked immunoassay (ELISA) [33], and liquid chromatography-tandem mass spectrometry (LC-MS/MS) [34]. However, these approaches have drawbacks such as expensive costs, complex operation, low detection sensitivity, and the need for complex pre-concentration during sample preparation and analysis [35–38]. Consequently, the advancement of uncomplicated, rapid, specific and high-throughput detection techniques [39].

Biosensors, utilizing enzymes, antibodies, aptamers, or other biomolecules, have recently enabled the detection of a wide array of viruses and pathogens [40–44]. The demand for these sensors is growing, with a particular need for biosensors that are simple, cost-effective, highly sensitive, and selective [45, 46]. Detection by these sensors involves two successive processes: biometrics, which detects specific biological responses for the selective identification of target substances, and signal transduction, which converts these responses into readable signals [47–50]. There is a broad variety of biosensors, with electrochemical and optical types being especially prevalent due to their wide usage and effectiveness [51–54]. Aptamers, which are short DNA or RNA sequences identified through the systematic evolution of ligands by exponential enrichment process (SELEX), attach to particular molecules with strong binding capacity, forming the basis for many DNA-based detection methods [55–61]. Here, we review the application of DNA sensors for aflatoxin B1 detection, with focus on electrochemistry-based biosensors, optics-based biosensors, and electrochemical luminescence sensors.

## Electrochemistry-based DNA biosensors

Currently, the detection of target substances using electrochemical biosensors is widely studied and utilized across multiple fields, including medical diagnosis, food safety monitoring, chemical hazard assessment, and new drug screening [62–65]. An electrochemical DNA sensor is a device that utilizes immobilized DNA as a molecular recognition component on the electrode surface (Fig. 1). Electrical signals are produced directly at the electrode without the need for additional signal transduction devices, enabling a cohesive link with downstream electronics and data handling [66]. These sensors offer advantages such as

low cost, fast response speed, easy miniaturization, high sensitivity, and high selectivity, making them suitable for the detection of aflatoxin B1 [67–69].

This article focuses on DNA sensor platforms used for the detection of aflatoxin B1 through electrochemical techniques, including electrochemical impedance spectroscopy (EIS), amperometry, voltammetry, and potentiometry. Table 1 summarizes the published research on the advancement of electrochemical DNA sensors for aflatoxin B1 detection.

## Electrochemical impedance spectroscopy-based DNA sensors

The efficacy of electrochemical detection depends both on reliable biometric components and on sensitive transduction approaches. Electrochemical impedance spectroscopy (EIS) enhances the applicability of conventional electrochemical techniques by boosting signal sensitivity and achieving low detection limits [70]. An EIS-based DNA sensor transduces electrical signals through alterations in the interfacial properties between electrodes and electrolytes, caused by DNA hybridization, conformational shifts, or DNA damage. This approach serves as a powerful and sensitive tool for characterizing surface-modified electrodes [71]. Such sensors offer unique advantages in the detection process, including the capacity to distinguish surface binding occurrences from the resistance of the solution, simplifying the measurement of signal intensity, and minimal or no need for markers or indicators, thereby reducing detection time and complexity [72–74].

Liu et al. [75] developed an innovative electrochemical aptamer sensor with signal-off functionality that uses a click-polymerization strategy to amplify the signal for swift and accurate detection of aflatoxin B1 at a low cost. In this sensor, a thioaptamer was self-assembled onto a gold electrode surface through an Au–S bond, while the polymer P (BFPA-DES) (bis-ferrocenoyl propargylamide-4, 4'-diazido-2, 2'-stilbenedisulfonic acid disodium salt tetrahydrate) served as the signal probe to enhance the electrical signal. The c-DNA functionalized by the azide-compound was hybridized with the aptamer to form dsDNA through base pairing. When aflatoxin B1 was present, the aptamer exhibited a stronger affinity for aflatoxin B1 than for c-DNA, leading to aflatoxin B1 binding to the aptamer and displacing the complementary DNA. This interaction altered the electrochemical impedance characterization, reducing resistance and modifying the current signal. The sensor offered a linear detection range from 2 fg/mL to 2 ng/mL, with a limit of detection (LOD) of 0.804 fg/mL (Fig. 2).
**Fig. 1** Basic principles of electrochemical DNA sensors. A: general steps for electrochemical DNA sensing. In the electrochemical DNA sensor, the analyte passes through the sensing interface, which is functionalized with an immobilized DNA aptamer, generating a responsive electrical signal. B: main sensing processes of two electrochemical DNA sensors. The surface of sensor has the probe DNA immobilized on it, and after the addition of the target DNA, the probe DNA is specifically bound to the target DNA, resulting in the change of the current signal. DNA: deoxyribonucleic acid

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## Amperometric-based DNA sensors

The amperometric technique is an electrochemical sensing that involves maintaining a steady potential at the working electrode to stimulate the analyte's redox processes on the electrode. The output current generated during the redox process is proportional to the concentration of the analyte [76]. In practical applications, amperometry is widely used in electrochemical sensor detection due to its accuracy, selectivity, high sensitivity, and minimal requirements for the composition and concentration range of the detected samples [77–79].

Mavrikou et al. [80] developed a Vero cell-based cellular biosensor that incorporates a membrane with an anti-AFB1 antibody as a biometric element, which interacts with aflatoxin B1 molecules at gold nanoparticles/screen-printed electrodes (SPE) within a three-electrode setup. The real-time current change was measured by chronoamperometry, to detect aflatoxin B1. The biosensor systems would be integrated with membrane-engineered cells. Once coated with poly (L-lysine), either typical Vero cells or Vero-anti-AFB1 cells were nurtured on the electrode's surface. After adding varying concentrations of aflatoxin B1 at 0.15, 0.5, 1.5, 3,

10, 15, and 30 ng/mL, the Vero cells captured aflatoxin B1 molecules, causing a certain degree of change in the current signal. The approach demonstrated exceptional sensitivity, with a limit of detection (LOD) of 0.5 ng/mL for Vero cells.

## Voltammetric-based DNA sensors

Voltammetry is the most widely employed method in electrochemical detection, known for its high accuracy, precision, sensitivity, and selectivity. Types of voltammetry include square wave voltammetry (SWV), differential pulse voltammetry (DPV), cyclic voltammetry (CV), and linear sweep voltammetry (LSV). Among these methods, square wave voltammetry and differential pulse voltammetry exhibit optimal detection sensitivity [81, 82]. Cyclic voltammetry and differential pulse voltammetry, in particular, are recognized as the most optimal, straightforward, cost-effective, and widely used techniques for assessing food quality. Their primary advantage is their ability to detect low concentrations of various analytes in a single step [83]. Voltammetric-based DNA sensors can detect DNA hybridization events by monitoring changes
<caption>**Table 1** Electrochemical DNA sensors for aflatoxin B1 detection. P (BFPA-DES): polymer bis-ferrocenoyl propargylamide-4,4'-diazido-2, 2'-stilbenedisulfonic acid disodium salt tetrahydrate.</caption>

ssDNA: single-stranded deoxyribonucleic acid. AFB1: aflatoxin B1. Tris-HCl: tris(hydroxymethyl)aminomethane hydrochloride

<table><thead><tr><th>Method</th><th>Assay strategy</th><th>Nanomaterial</th><th>Sensor detection buffer</th><th>Recognition motif</th><th>Linear range</th><th>Limit of detection</th><th>References</th></tr></thead><tbody><tr><td>Electrochemical impedance spectroscopy</td><td>Click aggregation P(BFPA-DES) to cause signal amplification strategy</td><td>Polymer Bis-Ferrocenoyl Propargylamide-4,4'-diazido-2,2'-stilbenedisulfonic acid disodium salt tetrahydrate</td><td>Phosphate buffered saline</td><td>ssDNA</td><td>2 fg/mL-2 ng/mL</td><td>0.804 fg/mL</td><td>[74]</td></tr><tr><td>Amperometric</td><td>Based on Vero cell electrical insertion and osmotic insertion</td><td>Gold nanoparticles</td><td>Phosphate buffered saline</td><td>Anti-AFB1</td><td>—</td><td>0.5 ng/mL</td><td>[79]</td></tr><tr><td>Voltammetric</td><td>Rolling circle amplification</td><td>Gold nanoparticles bonded to cerium-metal-organic frameworks</td><td>Phosphate buffered saline</td><td>ssDNA</td><td>0.024-100 ng/mL</td><td>1.48 pg/mL</td><td>[85]</td></tr><tr><td>Voltammetric</td><td>Polyadenine-based tetrahedral DNA nanostructure</td><td>Tetrahedral DNA nanostructure</td><td>Tris-HCl</td><td>ssDNA</td><td>1 fg/mL-1 ng/mL</td><td>0.33 fg/mL</td><td>[86]</td></tr><tr><td>Voltammetric</td><td>Dual function strategy based on tetrahedral DNA nanostructure</td><td>Au</td><td>Phosphate buffered saline</td><td>ssDNA</td><td>0.05-360 ng/mL</td><td>3.5 pg/mL</td><td>[87]</td></tr><tr><td>Potentiometric</td><td>The transfer spectrum is changed based on Au-8-pyrrole-dimercaptan</td><td>Au-8-pyrrole-dimercaptan</td><td>Phosphate buffered saline</td><td>Au-8-pyrrole-dimercaptan detecting element</td><td>—</td><td>—</td><td>[97]</td></tr></tbody></table>

in electrochemical parameters or the redox activity of electroactive markers, following the addition of a sample containing the target DNA [84].

In recent years, black phosphorus nanosheets (BPNs) have emerged as a research topic in the fields of medicine and chemistry due to their distinctive electrical and optical properties, as well as favorable biocompatibility. Black phosphorus nanosheets possess high electron mobility and considerable specific surface area, which makes them valuable for the detection of aflatoxin B1 [85]. Liao et al. [86] developed an innovative sensor based on DNA nanosheets. The hybridization between prism DNA and a primer probe was used to trigger rolling circle amplification (RCA) on the electrode surface, while a complementary chain modified by AuNPs@Ce-MOF combined with the amplified chain to produce DPV response signals (Fig. 3). The sensor's DPV response current demonstrated a strong linear correlation with the log of aflatoxin concentrations from 0.024 to 100 ng/mL, achieving a limit of detection down to 1.48 pg/mL (S/N of 3).

The tetrahedral DNA nanostructure (TDN) has high 3D mechanical rigidity and excellent programmability, but it presents certain limitations in the performance of electrochemical biosensors. To enhance the performance of the sensor, Wang et al. [87] innovative tetrahedral DNA nanostructure (ATDN) based on polyadenine for fabricating electrochemical aptamer sensors. The ATDN was constructed from four well-designed DNA single strands, with three polyadenine (PolyA) fragments at its base and five vertices containing aflatoxin B1 aptamers. The 5' signal probe with ferrocene can be fixed in place by aptamers to generate a substantial electrochemical response. After adding aflatoxin B1, the aptamer captures it and releases the signal probes, resulting in a drop in the electrical signal and enabling efficient detection of aflatoxin B1; it demonstrated a linear response across the range of 1 fg/mL to 1 ng/mL, with a recovery rate ranging from 95.2 to 98.9%.

Li et al. [88] successfully exploited a TDN-based dual-functional biosensor for the concurrent identification of aflatoxin B1 and ochratoxin A (OTA). AFB1@TDNs and
<figure><img src="image_5.png" /><figcaption>**Fig. 2** The electrochemical aptamer sensor for aflatoxin B1 (AFB1) utilizing click polymerization. The aptamer DNA was immobilized on the sensor surface using Au–S bonds. MCH was then added to block unbound non-specific sites, followed by the introduction of azide-functionalized c-DNA, which hybridized with the aptamer DNA to form double-stranded DNA. P (BFPA-DES) was subsequently loaded onto the double-stranded DNA through a click chemistry with the azide group, resulting in an increase in the electrical</figcaption></figure>

signal. Upon the addition of aflatoxin B1, it displaced the c-DNA on the aptamer, leading to the detachment of P (BFPA-DES) and a decrease in the electrical signal. HS: sulfhydryl. BFPA: bis-ferrocenoylpropargylamide. DES: 4, 4'-diazido-2, 2'-stilbenedisulfonic acid disodium salt tetrahydrate. MCH: 6-mercapto-1-hexanol. DNA: Deoxyribonucleic acid. c-DNA: complementary deoxyribonucleic acid

OTA@TDNs were fixed on the gold electrode surface via an Au–S bond, and Fc@DNA could be partially hybridized with AFB1@TDNs to form a finely separable nano-polymer. When aflatoxin B1 was added, the polymer disintegrated, especially at low temperatures, and recombined with AFB1@TDNs in place of Fc@DNA. Due to the disconnection of Fc@DNA, the electrical signal attenuated significantly, enabling precise detection of aflatoxin B1. At the ideal conditions, the peak current exhibited a linear response to aflatoxin B1 concentrations ranging from 0.05 to 360 ng/mL, with a limit of detection (LOD) of 3.5 pg/mL. The current of ferrocene and the concentration of aflatoxin B1 showed a linear relationship between 0.05 and 360 ng/mL, with a detection limit of 3.5 pg/mL.

## Potentiometric-based DNA sensors

Potentiometric biosensors are among the oldest biosensing platforms, commonly used to determine ion concentrations by measuring the charge produced by the analyte [89]. The current between the electrodes is minimal, and the build-up of electrical charge potential at the working electrode is primarily assessed and contrasted with that of the reference electrode within the electrochemical cell [90]. A characteristic element of potential sensor design is the induction layer, where a potential difference is generated at the sensor-solution interface [91]. This sensor stands out from

other analytical tools due to its excellent analytical properties, affordable and user-friendly, and resistance to interference from color and turbidity [92–94]. Specific DNA can be introduced into potentiometric sensors as aptamers, allowing the sensor to achieve excellent specificity and selectivity in detecting the target analyte [95, 96]. Recent studies have shown that the working principle of potential transduction is contingent upon the potential disparity between the sensing electrode and the reference electrode. When zero or negligible flows through the electrode, the potential difference accumulates during the recognition process in the electrochemical cell. Conductance transduction in micro/nano electronic devices, such as field-effect transistors could directly monitor changes in electrode conductance caused by DNA, hybridization, or complementary antibody-antigen equivalence binding [97].

Mo et al. [98] studied gold-8-pyrrole-dithiol molecular dots (Au-8PyDT) for their application as an electrochemical single-molecule sensor (SMS) to detect aflatoxin B1. This single-molecule junction forming field-effect transistor sensor, also known as MolFET, could adsorb aflatoxin B1 in the Au-8PyDT molecular channel via hydrogen bonding, modifying the space in the transmission region and significantly changing the single-molecule sensors transmission spectrum. This interaction led to µA-level current changes in SMS. The sensing performance was significantly improved by applying an appropriate gate voltage, which reduced the
<figure><img src="image_6.png" /><figcaption>**Fig. 3** The aptamer sensor for aflatoxin B1 detection based on a DNA nanosheet structure. The constructed DNA nanoprism sensor hybridizes with the AFB1 aptamer through complementary binding. During the detection process, AFB1 specifically binds to the aptamer, causing the aptamer to dissociate from the DNA strand. Subsequently, the DNA1 probe, which is complementary to the DNA prism, hybridizes and binds to the electrode surface. Following this, the introduction of padlock probe, primer probe, and phi29 DNA polymerase initiates the rolling circle amplification (RCA) reaction on the electrode surface. Finally, the addition of DNA2 modified with AuNPs@Ce-MOF, which is complementary to DNA1, enables the immobilization</figcaption></figure>

of AuNPs@Ce-MOF onto the electrode, thereby achieving a significant enhancement in the current response signal. In contrast, without AFB1 participating in the process, DNA1 cannot bind to the electrode and the current response signal is not enhanced. AFB1: aflatoxin B1. AuNPs: gold nanoparticles. MOF: metal-organic framework. DPV: differential pulse voltammetry. Phi 29: Phi 29 deoxyribo-nucleic acid polymerase. DNA1 chain (5' to 3'): TGAGGTAGTATT TTTTTTTTTTTTTTTTTT. DNA2 chain (5' to 3'): SH-C6-TATCTC TATCTC. Primer probe chain (5' to 3'): TACTACCTCAATCCCTAT AAATACCCTAAC. Padlock probe chain (5' to 3'): p-TTATAGGGT ATCTCT TCTCTTAGGGTAT

current from the µA-level to the nA-level, thus maintaining stability. By analyzing the adjusted potential difference curve, stable detection of aflatoxin B1 was achieved.

## Optical DNA biosensors

An optical DNA biosensor identifies target analytes by monitoring alterations in optical responses, including refractive index, color, and fluorescence intensity, which are caused by the interaction of the analyte with the detection reagent [99–101]. Optical biosensors are advantageous as they offer essential information into the sample, encompassing kinetic properties, concentration levels, and molecular affinities, all while minimizing interference from electrical or magnetic fields [102]. Additionally, optical biosensing has been widely applied in biosensors due to its highly sensitive and non-destructive detection capabilities [103, 104]. The optical DNA sensor operates on the principle of

DNA hybridization, where aflatoxin B1 binds to a specific DNA aptamer sequence, resulting in a measurable change in fluorescence signal or absorbance. This section focuses on the detection of aflatoxin B1 using widely applied optical sensing methods, such as the colorimetric method, surface plasmon resonance (SPR), photonic crystal, fluorescence method, and electrochemiluminescence. Table 2 summarizes the advancements in optical DNA sensors for aflatoxin B1 detection. Based on the data in Table 2, the limit of detection and linear range of the optical sensor detection AFB1 methods are summarized and arranged, respectively (Figs. 4, 5).

## Colorimetric-based optical DNA sensors

Colorimetric sensors allow for the detection of substances by utilizing nano-enzymes or materials mimicking nano-enzymes to drive chemical transformations that produce color differences [105]. These sensors are favored for their cost-effectiveness, sensitivity, and specificity, with results
<caption>**Table 2** Optical DNA sensors for aflatoxin B1 detection. TAE: tris-acetate-EDTA. DNA: deoxyribonucleic acid. ssDNA: single-stranded deoxyribonucleic acid. dsDNA: double-stranded deoxyri-</caption>

bonucleic acid. AFB1: Aflatoxin B1. DTS: DNA tetrahedral scaffold. Tris–HCl: Tris (hydroxymethyl) aminomethane hydrochloride

<table><thead><tr><th>Method</th><th>Assay strategy</th><th>Nanomaterial</th><th>Sensor detection buffer</th><th>Recognition motif</th><th>Linear range</th><th>Limit of detection</th><th>References</th></tr></thead><tbody><tr><td>Colorimetric</td><td>Introduction of hybrid chain reaction (HCR) based on photonic crystal-DNA hydrogels</td><td>Photonic crystals -DNA hydrogel</td><td>The TAE/Mg<sup>2+</sup> buffer</td><td>Lap DNA</td><td>20 nM–2 µM</td><td>–</td><td>[109]</td></tr><tr><td>Colorimetric</td><td>Amplifying the signal based on catalytic DNA circuits</td><td>Gold nanoparticles</td><td>10 mM Tris-HCl, 120 mM NaCl, 5 mM KCl, 5 mM MgCl<sub>2</sub>, pH 7.4</td><td>ssDNA</td><td>10 pM–µM</td><td>2 pM</td><td>[114]</td></tr><tr><td>Colorimetric</td><td>The chimeric aptamer is bound to gold nanoparticles</td><td>Gold nanoparticles</td><td>10 mM Hepes, 120 mM NaCl, 5 mM MgCl<sub>2</sub></td><td>dsDNA</td><td>5–5120 nM</td><td>1.88 nM</td><td>[118]</td></tr><tr><td>Colorimetric /Fluorescence</td><td>Introduction of Nb<sub>2</sub>C and Mn<sup>2+</sup> enhances the signal</td><td>Niobium carbide</td><td>Tris-HCl</td><td>ssDNA</td><td>1–800 ng/ mL/0.1–450 ng/mL</td><td>0.0950 ng/mL</td><td>[119]</td></tr><tr><td>Surface plasmon resonance</td><td>AFB1 competes with streptavidin for biotin-nizedaptamers</td><td>–</td><td>Phosphate Buffered Saline</td><td>ssDNA</td><td>1.5–50 ng/mL</td><td>0.19 ng/mL</td><td>[128]</td></tr><tr><td>Fluorescence</td><td>Enzyme-free labeling fluorescence assay</td><td>Graphene</td><td>Phosphate Buffered Saline</td><td>dsDNA</td><td>5–50 nM</td><td>0.76 nM</td><td>[137]</td></tr><tr><td>Fluorescence</td><td>Signal switch fluorescence detection based on DTS</td><td>–</td><td>Tris-HCl; 20 mM Tris, 50 mM MgCl<sub>2</sub>, pH 8.0</td><td>Hairpin DNA</td><td>0.05–100 ng/mL</td><td>0.01 ng/mL</td><td>[140]</td></tr><tr><td>Electrochemiluminescence</td><td>The combination of DTS and DNA walkers amplifies the signal efficiently</td><td>Bis (2,2'-bipyridine) (5-amino-1,10-phenanthroline) ruthenium(II)</td><td>Acetate buffer</td><td>dsDNA</td><td>1.0 fg/ mL–10 ng/mL</td><td>0.58 fg/mL</td><td>[152]</td></tr><tr><td>Electrochemiluminescence</td><td>Two-channel zirconium fiber optic inspection</td><td>Zr-Metal-organometallic frame</td><td>Tris-HCl</td><td>ssDNA</td><td>0.001–30 ng/mL</td><td>0.6 pg/mL</td><td>[156]</td></tr></tbody></table>

often easily determined by simple visual observation. Over the past decade, colorimetric sensors have been extensively employed for detecting a range of compounds, organic contaminants, heavy metal ions, and biological molecules, making them valuable in fields such as food safety inspection, clinical disease diagnosis, and environmental pollution control [106–108]. When combined with DNA functionalization, the effectiveness of colorimetric DNA sensors is further enhanced.

Among optical colorimetric detectors, photonic crystals have garnered considerable interest because of their distinctive capacity to control the movement of light. Photonic

crystals-based biosensors offer several advantages over other competing biosensing technologies, including cost-effective manufacturing and short analysis times, making them useful for the detection of aflatoxin B1 [109, 110]. Chen et al. [111] first developed a photonic crystal-DNA hydrogel sensor that induced optical signal changes through a hybrid chain reaction (HCR). This sensor was equipped with a specific DNA sequence aptamer for aflatoxin B1, enabling highly selective and sensitive detection while also amplifying the signal. The interaction at a specific binding site with hairpin DNA triggered a hybridization chain reaction that elongated the strand of DNA linker, resulting in the enlargement of the 2D
<figure><img src="image_7.png" /><figcaption>**Fig. 4** Limits of detection of DNA biosensors based on Table 2 data. $[Ru(bpy)_2(5-NH_2-phen)]^{2+}$: Bis (2, 2'-bipyridine) (5-amino-1,10-phenanthroline) ruthenium (II)</figcaption></figure>

photonic crystals NA hydrogel. The hydrogel demonstrated a favorable reaction to aflatoxin B1 concentrations between 20 nM and 2 µM and displayed superior selective detection capabilities.

Colorimetric biosensors harnessing gold nanoparticles to relay signals are becoming more popular owing to their uncomplicated nature, ease of employment, miniaturized form, and readiness for field examinations [112–115]. Chen et al. [116] invented an enzyme-free biosensor with a catalytic DNA ring as the core, specifically for aflatoxin B1 amplification detection. The sensor utilized streptavidin non-functional gold nanoparticles as signal indicators and adopted cascade signal amplification. By using streptavidin-modified gold nanoparticles to act as a signal transducer, the target solution was easily identified by directly observable red-to-blue variations. In the absence of aflatoxin B1, streptavidin-functionalized gold nanoparticles dispersed in the ultraviolet-visible spectrum, with narrow absorption peaks occurring at 520 nm. Conversely, in the presence of aflatoxin B1 was present, there was a broad absorption spectrum was observed between 650 and 700 nm, while absorbance at 520 nm lowered. The colorimetric biosensor

demonstrated exceptional sensitivity for aflatoxin B1 detection. The calibration curve showed a linear response from 10 pM to 1 µM, with a detection limit (LOD) of 2 pM.

In the detection of aflatoxin B1 using DNA sensors, nucleic acid aptamers play a crucial role. The synergy between aptamers and nanomaterials enhances the high selectivity and reactivity of biosensors, offering a novel approach in the field and enabling convenient, prompt sensing, and identification of desired molecules [117–119]. Yang et al. [120] developed a gold nanoparticle (AuNPs) aptamer sensor to further verify the practical application of chimeric aptamers. Through engineering optimization of the initial anti-AFB1 aptamer, a chimeric aptamer was transformed to improve the detection of aflatoxin B1. This was achieved by combining the modified chimeric aptamer with the peroxidase-like activity of gold nanoparticles. The sensor exhibited high sensitivity and specificity, with a linear range for aflatoxin B1 in corn oil samples of 5–5120 nM and the limit of detection was 1.88 nM.

Kong et al. [121] engineered a two-function induction sensor and a two-track artificial neural network advanced testing framework for the exact quantification of aflatoxin
<figure><img src="image_8.png" /><figcaption>**Fig. 5** DNA biosensors linear range based on Table 2 data. [Ru(bpy)₂(5-NH₂-phen)]²⁺: Bis (2, 2'-bipyridine) (5-amino-1,10-phenanthroline) ruthenium (II)</figcaption></figure>

B1 in food samples. Surface-depressed niobium carbide MXene nanomaterials (Nb₂C-MNs) were first prepared, and introducing manganese ions (Mn²⁺) considerably amplified the adherence of Nb₂C to single-stranded deoxyribonucleic acid (ssDNA). In the presence of aflatoxin B1 within the detection system, the separation of ssDNA labeled with 6-carboxyfluorescein (ssDNA-FAM) and niobium carbide MXene nanomaterials occurred due to the high binding affinity of aflatoxin B1 for ssDNA, leading to an increased fluorescence response or a reduced colorimetric response. With the aforementioned principles as a foundation, a dual-modality detection approach for aflatoxin B1, involving both fluorescence and colorimetry, was accomplished (Fig. 6). The LOD was 0.0950 ng/mL under ideal conditions, and the operation was simple, rapid, and high selectivity, which enabled to satisfy the needs for prompt field analysis of aflatoxin B1.

**Surface plasmon resonance-based optical DNA sensors**

Surface plasmonic resonance (SPR) is a label-free detection method. When the incident light photon strikes the metal surface, it excites surface plasma, allowing for the detection of refractive index alterations in the medium adjacent to the metallic surface. Any variation in the refractive index affects the propagation speed of the surface plasmons [122]. Since its introduction in the early 1990s, SPR has become a powerful tool for studying interactions between biomolecules based on affinity binding analysis of multiple bonds [123]. When a DNA aptamer is used as a probe on the metal surface, the analyte solution comes into contact with the metal surface, enabling specific recognition between the DNA probe and the analyte. This interaction alters the surface refractive index, and the resulting signal change can
<figure><img src="image_9.png" /><figcaption>**Fig. 6** Principle of aflatoxin B1 (AFB1) detection using Nb₂C@ssDNA. The introduction of manganese ions (Mn²⁺) significantly enhances the affinity between Nb₂C and ssDNA, enabling the FAM-labeled ssDNA to be tightly immobilized on the sensor surface. Upon the addition of AFB1, the stronger affinity between AFB1 and ssDNA causes the dissociation of ssDNA-FAM from Nb₂C-MNs. This results</figcaption></figure>

in an increase in the fluorescence signal, while the colorimetric signal weakens upon the addition of H₂O₂ and TMB. Nb₂C-MNs: niobium carbide MXene nanomaterials. ssDNA: single-stranded deoxyribonucleic acid. FAM: 6-carboxyfluorescein. H₂O₂: hydrogen peroxide. TMB: 3, 3', 5, 5'-tetramethylbenzidine

be detected by an optical identifier [124–126]. As a relatively novel and straightforward analytical method, SPR has proven to be fast, sensitive, and label-free, with broad applications in food analysis and detection [127–129].

Wu et al. [130] created a surface plasmon resonance biosensor based on aptamer-functionalized for the identification of aflatoxin B1, effectively utilizing it for the immediate identification of aflatoxin B1 in vinegar samples. Upon immobilizing streptavidin onto the CM5 sensor chip via amine-coupled covalent bonding, the strong affinity of streptavidin for biotin facilitated the binding of the biotin-labeled adapter to streptavidin, thus allowing for its capture. By adding different concentrations of aflatoxin B1 to vinegar samples, changes in the refractive index response signal were observed on the CM5 sensor surface due to the strong binding interaction between the biotinized aptamer and aflatoxin B1. Furthermore, when the aflatoxin B1 injection ceased and the buffer solution was allowed to flow over the chip surface, the response signal swiftly reverted to its initial baseline level. The results showed that aflatoxin B1 bound to the aptamers in a fast-binding and fast-dissociation manner, implying that aflatoxin B1 could be released from the binding site by the buffer without requiring any regenerating agents, allowing for quick entry into the next cycle. This feature enabled rapid transition to the next cycle, demonstrating that the chip could be reused multiple times without compromising detection sensitivity, thus reducing ligand consumption and lowering costs. The biosensor

demonstrated a linear response between 1.5 and 50 ng/mL, and 0.19 ng/mL was the detection limit of the sensor.

## Fluorescence-based optical DNA sensors

Fluorescence sensors are of great significance and function in biological research and have gained increasing attention in recent years due to their advantages, including high throughput, straightforward operation, low sample volume, high sensitivity, and non-radioactivity [131]. The construction of such sensors could be centered around fluorescent proteins, probes, or amalgamated systems [132, 133]. The robust helical configuration of DNA allows fluorescence resonance energy transfer (FRET) to be transmitted over relatively long distances along the strand, rendering the DNA framework an ideal scaffold for these multi-chromophore sensors [134]. The combination of DNA and fluorescence technology has facilitated the progress of numerous biosensors for aflatoxin B1 detection, which are widely used in food detection [133].

Graphene-based DNA sensor technology exhibits excellent selectivity, real-time analysis capabilities, and ease of synthesis, demonstrating significant application potential in the detection of pathogens and infectious diseases [135]. The introduction of graphene oxide (GO) nanomaterials has further enhanced the sensitivity of DNA-based fluorescence sensors, offering excellent properties such as hydrophilicity, biocompatibility, and fluorescence quenching [136, 137]. Because graphene oxide has a strong ability
to adsorb single-chain parts and quench fluorescence, the background fluorescence is effectively reduced [138, 139]. Hu et al. [140] developed a novel fluorescence sensor combining graphene oxide with SYBR Green I dye, which could easily detect aflatoxin B1 without washing and separation steps. The model consisted of a DNA double-stranded structure (AFB1 aptamer trigger chain) and two hairpin probes H1 and H2. When aflatoxin B1 was present, it interacted with the aflatoxin B1 aptamer and released a trigger chain, which was then bound to the hairpin probe and could emit a strong fluorescence signal under the action of SYBR Green I and graphene oxide. Conversely, when aflatoxin B1 was not absent, graphene oxide could adsorb dsDNA and hairpin DNA with long sticky ends well, resulting in weak fluorescence.

Previously, the DNA tetrahedral scaffold (DTS) became popular for biosensing due to its stable structure, strong mechanical rigidity, high precision, clear spacing, and specific orientation [141, 142]. Ren et al. [143] developed a signal-switching fluorescence sensor based on a DNA self-assembled tetrahedron, which allowed for the concurrent detection of ochratoxin A and aflatoxin B1 through the on-off functions of molecular markers. The DNA tetrahedral scaffold was composed of four unique single-stranded DNA (ssDNA) S1, S2, S3, and S4 through a process of controlled denaturation and annealing. Cy3 and Cy5 served as a molecular marker attached to the extension of DNA strands S3 and S4, respectively, while the black hole quencher 2 (BHQ2) was located at the opposite end. Upon the assembly of the DNA tetrahedral scaffold, the fluorescence of Cy3 and Cy5 was effectively quenched by the FRET effect. Upon the addition of ochratoxin A and aflatoxin B1 aptamers, which are complementary to the hairpin structure, the hairpin opened, restoring the fluorescence signal. When ochratoxin A and aflatoxin B1 were present, they bound to their respective aptamers, preventing the aptamers from opening the hairpin structure and thereby suppressing the fluorescence signal once again. The detection range for ochratoxin A was linear from 0.01 to 100 ng/mL, with a detection limit of 0.005 ng/mL, while the detection range for aflatoxin B1 was 0.05–100 mL, with a LOD of 0.01 ng/mL.

## Electrochemiluminescence-based DNA sensors

Electrochemiluminescence is the process of producing light by oxidizing or reducing luminescent groups to transfer radiative charge between electrochemically generated free radicals [144]. Known for its broad dynamic range, high sensitivity, and rapid analytical capabilities, electrochemiluminescence is significantly influential across numerous fields such as biomolecular detection, food safety and drug analysis, clinical diagnosis, and environmental monitoring [145–149]. When combined with DNA

technology, electrochemiluminescence DNA sensors are particularly promising due to their wide dynamic range, high sensitivity, and controllability [150]. The detection method is often accompanied by electrochemiluminescence tags, such as $Ru(bpy)_3^{2+}$ [151–153].

In electrochemiluminescence biosensors, the effective fixation of DNA probes is crucial for enhancing sensor sensitivity [154]. Li et al. [155] integrated the excellent mechanical strength of DTS with the potent signal enhancement function of DNA walker to develop an efficient and easy-to-operate electrochemical luminescence detection platform, which is specially used for the high-sensitivity detection of aflatoxin B1. On the electrode surface, DTS was immobilized through Au–S linkages, and $Ru(bpy)_2(cphen)_{2+}$ was embedded in the dsDNA component of DTS to achieve the starting electrochemiluminescence emission. The electrochemiluminescence signal of $Ru(bpy)_2(cphen)_{2+}$ was effectively quenchable by introducing ferrocene labeled at the end of the substrate chain. Upon introducing aflatoxin B1, the aptamer selectively engaged with aflatoxin B1, triggering the activation of the 3D DNAzyme and the cleavage of the substrate scaffold by $Na^+$, which led to the detachment of ferrocene from the electrode and the resurgence of the $Ru(bpy)_2(cphen)_{2+}$ electroluminescent signal (Fig. 7). Therefore, by evaluating the increase in the electrochemiluminescence response of $Ru(bpy)_2(cphen)_{2+}$, ultra-selective identification of aflatoxin B1 was accomplished, featuring a detection range from 1.0 fg/mL to 10 ng/mL and a limit of detection (LOD) at 0.58 fg/mL.

A metal–organic framework (MOF) is a nano-structured material with a three-dimensional structure composed of metal ions as nodes and organic ligands as the skeleton. MOFs are characterized by a large specific surface area, numerous metal centers, and ease of modification [156, 157]. Among them, Zr-MOF can be used in the design of biosensors because of the strong affinity of zirconium segments to phosphoric acid groups in biomolecules [158]. Li et al. [159] developed a two-channel zirconium oxide optical fiber aptamer sensor for accurate detection of aflatoxin B1. In this sensor, Zr-MOF serve as a signal probe was conjugated to the aptamer and fixed to the electrode surface by cDNA base complementary pairing. The organic ligand $NH_2$-BDC exhibited excellent electrochemical signal and fluorescence signals. Upon detection of aflatoxin B1, the aptamer selectively bound to aflatoxin B1, leading to the detachment of the Zr-MOF/aptamer probe and its entry into the solution, which caused a reduction in the electrochemical response and an increase in fluorescence intensity. The linear detection range for the electrochemical signal was 0.001~30 ng/mL, with an LOD of 0.6 pg/mL, while the LOD of the fluorescence signal was 0.8 pg/mL.
<figure><img src="image_10.png" /><figcaption>**Fig. 7** Mechanism of aflatoxin B1 (AFB1) detection based on DTS surface 3D DNAzyme walker. The Ru complex was embedded in the dsDNA framework of DTS, and the initial electrochemiluminescence signal was obtained. The electrochemiluminescence signal was suppressed by the introduction of labeled ferrocene. Aflatoxin B1, with</figcaption></figure>

the assistance of Na⁺, could bind to the aptamer, cleave the substrate scaffold, release ferrocene, and restore the electrochemiluminescence signal. DTS: DNA tetrahedral scaffold. SH: sulfhydryl. 3D: three-dimensional. Ru complex: Ru(bpy)₂(cphaphen)²⁺. dsDNA: double-stranded deoxyribonucleic acid

## Other methods of detection

In addition to DNA-based detection methods, other detection methods are also used to detect aflatoxin B1, for instance, enzyme-linked immunosorbent assay (ELISA) [160, 161], lateral flow immunochromatography assay (LFIA) [162], radioimmunoassay [163], ultra-high performance liquid chromatography-fluorescence detection [164, 165], competitive fluorescence microsphere immunoassay assay (CFIA) [166], and chemiluminescence immunoassay (CLIA) [167]. Table 3 summarizes the research progress of various methods for aflatoxin B1 detection.

Liquid crystals (LC) exhibit high birefringence and significant sensitivity to surface interactions at the water/LC interface. Cheng et al. [168] developed 4-cyano-4'-amyl biphenyl droplet (5CB<sub>SADrop</sub>) anchored on the upper surface of DMOAP (N, N-dimethyl-N-octadecyl (3-aminopropyl) trimethoxysilyl chloride) coated glass via solvent evaporation method, enabling the quick and sensitive detection of aflatoxin B1 based on the droplet conditions. When an aqueous solution of cetyltrimethylammonium bromide (CTAB) was dropped on 5CB<sub>SADrop</sub>, the droplet formed a dark cross pattern under the spatial conformation polarized optical microscope. After the addition of aflatoxin B1 aptamer,

the aptamer, and CTAB intertwined to form a mixture that escaped from the droplet, resulting in a four-leaf clover configuration in the radiation image. In the presence of aflatoxin B1, the toxin's interaction with the aptamer resulted in the detachment of the CTAB-AFB1 aptamer assembly. This allowed CTAB to return to the 5CB<sub>SADrop</sub>, restoring the dark cross configuration. The detection limit reached 10 pg/mL.

The suspension array technology based on cellular microbeads has garnered significant interest in recent decades due to its outstanding benefits such as strong flexibility, good repeatability, speed, rapidity, excellent specificity, and low cost [169]. Liu et al. [170] first established an environmentally friendly and trustworthy cytometric microbead magnetic suspension array (CBMSA) project and demonstrated its practicability and effectiveness in identifying minute concentrations of aflatoxin B1 in genuine lotus seed specimens. Utilizing the biotin-avidin interaction, biotinylated bovine serum albumin associated with aflatoxin B1 (BSA-AFB1, antigen) was affixed to the streptavidin-coated microbeads. Following magnetic separation and washing, the antigen-decorated microbead complex was acquired. When the primary antibody (PriAbs) of the mouse monoclonal antibody (McAbs) of aflatoxin B1 was added to the aflatoxin B1 standard solution sample, the aflatoxin B1 specimen and
<table><caption>**Table 3** Other methods to detect aflatoxin B1. ssDNA: single-stranded deoxyribonucleic acid</caption><thead><tr><th>Method</th><th>Assay strategy</th><th>Nanomaterial</th><th>Sensor detection buffer</th><th>Recognition motif</th><th>Linear range</th><th>Limit of detection</th><th>References</th></tr></thead><tbody><tr><td>Liquid crystals</td><td>Rapid detection based on droplet conditions</td><td>4-cyano-4'-amyl biphenyl droplet</td><td>Phosphate buffered saline</td><td>ssDNA</td><td>—</td><td>10 pg/mL</td><td>[165]</td></tr><tr><td>Cytometricmicrobead magnetic suspension array</td><td>Sensitive detection based on cytometric-microbead magnetic suspension array</td><td>Microbeads</td><td>Phosphate buffered saline</td><td>Primary antibody</td><td>—</td><td>7.8125 pg/kg</td><td>[167]</td></tr><tr><td>Lateral flow immunoassay</td><td>Luminescent organic compound-based microarray lateral flow immunoassay</td><td>Luminescent organic compound</td><td>Phosphate buffered saline</td><td>Goat anti-mouse</td><td>—</td><td>1.3 ppb</td><td>[168]</td></tr><tr><td>Field-effect transistor</td><td>Electrohydrodynamic-printed Ti<sub>3</sub>C<sub>2</sub>T<sub>x</sub>MXene-based field-effect transistor</td><td>Ti<sub>3</sub>C<sub>2</sub>T<sub>x</sub>MXene</td><td>Phosphate buffered saline</td><td>Anti-aflatoxin B1</td><td>0.7–20 ppb</td><td>5.689 ppb</td><td>[170]</td></tr></tbody></table>

the microbeads antigenic site on microbeads competed for PriAb binding in an antigen-antibody reaction. Following magnetic separation and subsequent washing treatment, the specific trapping probe MB-streptavidin-biotin-BSA-AFB1-MCAB was acquired. The signal reporting probe was integrated with the capture probe, to amplify the fluorescence signal. The suspended array fluorescence probe conjugate was acquired after treatment. In the end, a suspended array chip equipped with a fluorescent probe in each compartment was moved to a high-sensitivity flow cytometer for immediate fluorescence analysis. The LOD was 7.8125 pg/kg.

The lateral flow immunoassay (LFIA), synonymous with the immunochromatographic strip test, offers an alternative technique for detecting a range of analytes, boasting quick results, simplicity, point-of-care applicability, and low cost. Charlermrojet al. [171] invented a unique luminescent organic compound (LOC) as a signaling molecule with an extra-large Stokes shift. Based on the LOC, a microarray LFIA (μLFIA) was successfully established for simultaneous multi-detection of 5 mycotoxins: aflatoxin B1, T-2 toxin, zearalenone, fumonisin B1 and deoxynivalenol. The LOC, which possesses two aldehyde groups, was conjugated to the goat anti-mouse (GAM) antibody through a Schiff base formation and subsequent reduction reaction. In the absence of mycotoxins detected in the specimen, mycotoxin-directed antibodies would be unimpeded in their interaction with the M424-GAM antibody. In reverse, if the sample harbored any of the five mycotoxins, the respective antibodies would be occupied by these mycotoxins, which would result in a reduced or no signal. To examine the specificity of μLFIA in detecting one or multiple mycotoxins, a competitive assay was used to evaluate target and non-target mycotoxins with the best conditions that were previously identified. The minimum detectable levels of aflatoxin B1, T-2, ZON, FUMB1,

and DON were 1.3, 0.4, 0.9, 0.4, 0.4, and 0.5 ppb. The recovery rates for the above-mentioned toxins were between 70.7 and 119.5% for within-assay and between 80.4 and 124.8% for between-assay variations.

Field-effect transistors (FETs) offer impressive advantages, including ease of operation, miniaturization, and rapid response times [172]. During the FET sensor operation, external analytes easily cause alterations in the electrical conductivity of the sensing channel materials, which has good conditions for detecting low molecular weight aflatoxins. Siva et al. [173] report a field-effect transistors (FET) based on electrohydrodynamic-printed Ti<sub>3</sub>C<sub>2</sub>T<sub>x</sub>MXene as a viable biosensing solution for detecting aflatoxin B1. FETs were constructed using EHD printing technology on the terminal electrodes, which include the source and drain, and featured fluctuating layers of nanosheets. The anti-AFB1 antibodies were linked to the MXene surface for the creation of an AFB1 FET biosensor, and fixing a particular antibody to the FET reduced the channel's conductivity. After adding a certain concentration of aflatoxin B1, it bound to the AFB1 antibody on the surface, resulting in an immediate reduction in the drain current of the sensing chamber. The LOD was 5.689 ppb, with the optimal recovery of aflatoxin B1 from the samples reaching 98.4 ± 0.2%.

## Conclusion

We reviewed aptamer biosensors for efficient detection of aflatoxin B1, focusing on electrochemical, optical, and electrochemiluminescence-based methods. Some DNA biosensors incorporate excellent nanomaterials such as black phosphorus nanosheets, gold nanoparticles, and graphene oxide, which are widely used in optical and electrochemical
biosensor detection. These nanomaterials offer a large spe-
cific surface area, and high electron mobility. However, some
sensors also face challenges, such as negative effects on mass
and electron transport of functional and stable molecules, as
well as the tendency of nanomaterials to dissolve and aggre-
gate. The intricate nature of biological sample matrices often
introduces interference, potentially leading to false-positive
signals. While DNA-based sensors have achieved break-
throughs in reversibility and target-specific recognition, the
majority still suffer from inadequate response times and sen-
sitivity. Reported DNA sensors typically exhibit response
durations ranging from tens of minutes to over an hour,
highlighting a considerable sensitivity gap when compared
to modern PCR technology.

**Acknowledgements** This work was made with the support of National Natural Science Foundation of China (NSAF, No. U2230132), the open fund of Information Materials and Intelligent Sensing Laboratory of Anhui Province (Grant No. IMIS202209), Guiding Science and Technology Plan Project for Social Development in Zhenjiang City (FZ2022052), Zhenjiang Science and Technology Plan (Social development, SH2024098), the National Foreign Experts Program Project of China (H20240553), National Key R&D Program of China (2023YFC3503804), and Key Laboratory of Medicinal Animal and Plant Resources of Qinghai-Tibetan Plateau in Qinghai Province.

Declarations

**Conflict of interest** The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. EL and LG declare that they are Editors of Environmental Chemistry Letters.

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