Ultrasensitive Detection Research Articles

Highly Efficient and Rapid Surface-Enhanced Raman Spectroscopy Active Platform Based on Au Nanorods for Ultrasensitive Traumatic Brain Injury Detection

Highly Efficient and Rapid Surface-Enhanced Raman Spectroscopy Active Platform Based on Au Nanorods for Ultrasensitive Traumatic Brain Injury Detection

Ultrasensitive detection of remote acoustic vibrations at 300 m distance by optical feedback enhancement

Sensitive detection of remote vibrations at nanometer scale owns promising potential applications such as geological exploration, architecture, and public security. Nevertheless, how to detect remote vibration information with high sensitivity and anti-disturbance has become a major challenge. Reported current non-contact measurement methods are difficult to simultaneously possess characteristics of high light intensity sensitivity, long working distance, high vibration response sensitivity, and anti-disturbance of ambient light. Here, we propose a polarization-modulated laser frequency-shifted feedback interferometry method with the above characteristics, to obtain remote vibration information. The method can directly measure non-cooperative targets without the need for any cooperative markers. In each interference cycle, the energy as low as 2.3 photons can be effectively responded to, and the vibration amplitude sensitivity at 300 m can reach 0.72 nm/Hz1/2 at 1 kHz. This approach provides a strategy for the ultrasensitive detection of remote vibration that is immune to electromagnetic interference.

Construction of “ant-like tentacle” structure for ultra-sensitive detection of low-concentration ammonia through colorimetric fluorescent dual-signal gas-sensitive cotton fabric

Detection and monitoring of ammonia (NH3) are crucial in various industries, including plant safety management, food freshness testing, and water pollution control. Nevertheless, creating portable, low-cost, highly sensitive, and easily regenerated ppm-level NH3 sensors poses a significant challenge. In this investigation, an innovative “ant-like tentacle” fabrication strategy was proposed, and a colorimetric fluorescent dual-signal gas-sensitive cotton fabric (PAH-fabric) for NH3 detection was successfully prepared by conventional dyeing using suitable molecular-level photoacid (PAH) sensitive units. The visual recognition lower detection limit of the ultra-low is 1.09 ppm-level. PAH-fabric is not only straightforward, convenient, and cost-effective to prepare, but it can also be efficiently regenerated and recycled multiple times (maintaining excellent gas-sensitive performance even after 100 cycles) by strategically leveraging volatile acid fumigation. Detailed molecular reaction mechanisms involved in the NH3 response and PAH-fabric regeneration are elucidated. PAH-fabric, available either as a portable kit or an alarm system, offers a promising approach for ultra-low NH3 detection. The demonstrated “ant-like tentacle” fabrication strategy introduces numerous possibilities for designing and developing sensors with adjustable response thresholds, particularly those requiring high sensitivity.

Ingenious dual-circle DNA walker-mediated electrochemical biosensor for rapid and ultrasensitive detection of microRNA

In this work, an ingenious dual-circle DNA walker (DCDW) with pretty fast walking speed and high amplification efficiency was developed for rapid and ultrasensitive electrochemical detection of microRNA-221 (miRNA-221) related to liver cancer, combined with the toehold-mediated strand-displacement reactions (TSDRs). Impressively, compared with the traditional DNA walker, the DCDW with unique double-stranded interlocked DNA nanostructure not only possesses higher stability, flexibility, and anti-entanglement ability, but also enables more functional domain in a smaller area, thereby enhancing the local concentration, which can greatly improve the working efficiency. As a validation, the electrochemical biosensor realized rapid and ultrasensitive detection of miRNA-221 with a reaction time of 15 min and detection limit down to 1.9 aM, and had been applied in MHCC97L and HeLa cancer cell lysates, thus providing an innovative insight to design intelligent functional interlocked DNA walkers for ultimate application in the construction of biosensing platform and miRNA detection in biological sample.

Design, synthesis, solvatochromism, and colorimetric detection of amines based on isolated perylene bisimide anionic radical π-dimers

Some π-molecular radicals typically exist in solution as π-dimers with metastable characteristics. Therefore, attempting to improve the stability of radical π-dimers is a fascinating endeavor. Herein, a series of hydroxyl-functionalised isolated perylene bisimide anionic radical π-dimers were obtained by using intramolecular hydrogen bond assistance in a one-step process for the first time. It is also found that these radical π-dimers exhibit solvatochromism and acid-base discoloration, when the intermolecular hydrogen bonds are disrupted. Perylene bisimide anionic radical π-dimers exhibit unique properties such as solvatochromism and acid-base discoloration. These characteristics make them suitable for ultra-sensitive colorimetric and fluorescent dual-signal detection of amines. Among them the π-dimer 3 achievable the lowest detection limits of 0.13 ppb, 0.02 ppb and 0.049 ppb for primary, secondary and tertiary amines, respectively. Additionally, colorimetric films prepared using these radical π-dimers demonstrate high sensitivity, enabling detection of amines at ppb levels. We believe that this class of radical π-dimers could be develop into a new class of environmental stimuli-responsive materials.

A Triple Signal Amplification Strategy for Accurate and Ultrasensitive miRNA-21 Detection.

A triple signal amplification strategy was integrated with a built-in double electrode and external energy storage device to fabricate a novel self-powered biosensor for ultrasensitive detection of miRNA-21. Specifically, DNA tetrahedra and haripin2-glucose oxidase are modified on the surface of the biocathode and bioanode by catalytic hairpin assembly (CHA) to achieve dual signal amplification. Moreover, triple signal amplification is realized by including an external capacitor. Consequently, the as-constructed self-powered biosensor demonstrates a low detection limit of 0.06 fM toward the miRNA-21 assay within the range of 0.1 fM to 10 pM. This study presents a practical and sensitive approach to timely cancer detection.

Red-emitting copper nanoclusters for ultrasensitive and selective detection of creatinine and its application in portable smartphone-based paper strips and polymer thin film

In this work, a quantitative paper-based sensor integrated with smartphone-application was developed using poly(sodium 4-styrenesulfonate) stabilized copper nanoclusters (PSS-CuNCs) as a fluorescence probe to detect creatinine (CRN). The surface morphology and optical properties of PSS-CuNCs were verified using UV–vis absorption, zeta-potential, photoluminescence, high resolution transmission electron microscopy (HR-TEM) and Fourier-transform infrared (FI-IR) and X-ray photoelectron (XPS) spectroscopy. PSS-CuNCs exhibited strong red fluorescence with the emission peak centered at 654 nm upon excitation at 390 nm. PSS-CuNCs was also used to determine CRN because of their excellent fluorescence properties. The fluorescence of PSS-CuNCs was quenched significantly by adding different concentrations of CRN, which mainly originated from the formation of ground state complexation. The PSS-CuNCs probe showed high sensitivity with a low limit of detection (LOD) of 2.48 nM and high selectivity even in the presence of other interfering analytes. This assay was also used to measure CRN in real samples. Additionally, test paper-based assay has been developed to detect CRN using smartphones. The developed assay provides a reliable, cost-effective, sensitive and portable on-site method to measure CRN levels by utilizing fluorescence color changes. Incorporation of PSS-CuNCs into poly(vinyl alcohol) resulted in the formation of fluorescent transparent polymer thin films with high photostability.

Unlocking multi-mode sensing potential: Phosphorus-doped graphitic carbon nitride quantum dots for Ag+, ciprofloxacin, and riboflavin analysis in environment and food matrices

The simultaneous detection of multiple analytes through a single fluorescence sensor is highly attractive. In this study, phosphorus-doped graphitic carbon nitride quantum dots (P-CNQDs) were developed, achieving multi-mode sensing through three distinct response mechanisms. The preparation involved using melamine as the carbon and nitrogen source and ammonium dihydrogen phosphate as the phosphorus source. Uniform and narrowly distributed P-CNQDs were successfully synthesized through chemical oxidation and hydrothermal methods, with an average size of 2.4 nm. These unique P-CNQDs exhibited fluorescence quenching through photo-induced electron transfer (PET) in response to Ag+. Additionally, the formation of hydrogen bonds and coordination interactions between P-CNQDs-Ag+ and ciprofloxacin (CIP) led to a pronounced fluorescence response to CIP by the chelation enhanced fluorescence (CHEF) mechanism. Furthermore, leveraging the principle of fluorescence resonance energy transfer (FRET), P-CNQDs-CIP served as a ratio fluorescence sensor for riboflavin (RF), enabling ultra-sensitive detection of RF. The combination of PET, CHEF, and FRET response mechanisms successfully facilitated multi-mode sensing for Ag+, CIP, and RF. The detection ranges were 0.05–100 μM, 0.002–2 μM, and 0.05–60 μM, with corresponding lowest detection limits of 17.1 nM, 1.1 nM, and 29.2 nM, respectively. This versatile sensor has been effectively applied to real samples, including the detection of river water and vitamin B2 tablets.

A digital aptasensor for the ultrasensitive detection of T-2 toxin by using single molecule array

T-2 toxin is a low molecular weight, refractory mycotoxin, often contaminating cereal-based foods, and can cause detrimental effects on human health such as nausea, vomiting, and long-term immune suppression. Therefore, highly sensitive detection platform for T-2 toxin test at the molecular level is an urgent need for food safety and public health. In this work, a digital aptasensor is developed for the ultrasensitive detection of T-2 toxin by combining aptamer and single-molecule array technology. Initially, T-2 toxin competed with the biotin-labeled complementary sequence probe to bind the T-2 toxin aptamer. The free biotin-labeled probe was then hybridized with a capture probe modified on the magnetic beads to form a stable double-stranded structure. After combining with streptavidin-enzyme, the structure was used for fluorescence imaging analysis via single-molecule microwell arrays. By correlating the concentration of T-2 toxin with the number of fluorescent microtiter wells, the ultra-sensitive detection of the T-2 toxin was realized. The developed digital aptasensor has demonstrated remarkable sensitivity toward T-2 toxin, with a detection range between 0.16 and 300 pM (approximately 0.075–140 pg/mL) and a detection limit of 44.5 fM (approximately 20.8 fg/mL). Additionally, 87.0 %-103.1 % recoveries were obtained from spiked samples in corn. This work offers a highly effective approach to detecting trace amounts of T-2 toxin, which has the potential to be used for enhancing food safety protocols, providing a powerful tool for industries and regulatory agencies to protect public health.

CuS hollow nanocages with rough surface to enhanced photothermal property for ultra-sensitive detection of furazolidone metabolites

Signal tracers play a key role in the lateral flow immunoassay (LFIA) for improving sensitivity and signal output capability. Photothermal signal has sparked significant excitement in designing high-performance LFIAs. Herein, CuS hollow nanocages (CuS HNCs) were synthesized for building a dual signal LFIA. CuS HNCs show excellent photothermal conversion efficiency due to their unique morphology and superior localized surface plasmon resonance (LSPR) effect. Meanwhile, the hollow-cage structure endows CuS HNCs with high light absorption and suitable colorimetric property. Furazolidone (FZD) metabolite was used as a model target in the developed LFIA to demonstrate its superior comprehensive performances brought by CuS HNCs. The limit of detection is as low as 0.099 ng mL−1 (colorimetric mode) and 0.075ng mL−1 (photothermal mode), respectively. Furthermore, standard recovery test in honey and shrimp samples was performed and the recovery rates ranged from 86.33 % to 116.71 %. This study not only introduces a new type of nanolabel characterized by an abundance of high-energy hot carriers and pronounced thermal effects, enabling the development of exceptionally sensitive immunosensor platforms but also extends applications of hollow plasmonic photothermal material in the point-of-care (POC) diagnostic field.

Disposable Zirconium trisulfide-Reduced graphene oxide modified conducting thread based electrochemical biosensor for lung cancer diagnosis

Flexible technology in sensors have received much attention in monitoring of human health through various physiological indicators. Thus, it drawn a lot of interest in the development of flexible substrate for the diagnosis of various diseases via analysis of analytes. Present work focusses on the development of ecofriendly, portable, flexible, conducting thread (Th) and used as smart substrate for fabrication of biosensor towards ultrasensitive detection of the lung cancer biomarker (cytoskeleton-associated protein 4; CKAP4). The zirconium trisulfide-reduced graphene oxide nanocomposite and poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate) (PEDOT:PSS) modified cotton thread based biosensor was fabricated via dip coating method. Next, successive immobilization of monoclonal antibodies of CKAP4 (anti-CKAP4) and bovine serum albumin (BSA) was performed via drop cast approach using fabricated electrode [nZrS3@rGO/PEDOT:PSS/Th]. The response of fabricated electrode (BSA/anti-CKAP4/ZrS3@rGO/PEDOT:PSS/Th) was recorded electrochemically versus CKAP4 concentration via chronoamperometry (CA). The results showed wider linear detection range of 6.25-800 pg mL−1, excellent sensitivity of 85.2 µA[log(pg mL−1)]-1cm−2 with good stability up to 42 days. The response of fabricated biosensor was supported by investigating response of CKAP4 biomarker present in patients of lung cancer (concentration as determined through enzyme-linked immunosorbent assay) and obtained results exhibited excellent correlation with that of standard samples.

Noninvasive Prenatal Genetic Screening of Cell-Free Fetal DNA for Early Prediction of β-Thalassemia Using Fiber Optic Nanogold-Linked Sorbent Assay.

β-Thalassemia is a prevalent type of severe inherited chronic anemia, primarily identified in developing countries. The identification of single nucleotide polymorphisms (SNPs) plays a vital role in the early diagnosis of genetic diseases. Here, we reported the development of an amplification-free fiber optic nanogold-linked sorbent assay method using a fiber optic particle plasmon resonance (FOPPR) biosensor for rapid and ultrasensitive detection of SNPs. Herein, MutS protein was selected as the biorecognition capture probe and immobilized on the sensing region to capture the target mutant DNA, which was hybridized with a single-base mismatched single-stranded DNA labeled by a gold nanoparticle (AuNP). The AuNP acts as a signaling agent to be detected by the FOPPR biosensor when it is bound on the fiber core surface. The method effectively differentiates mismatched double-stranded DNA by MutS protein from perfectly matched/complementary dsDNA. It exhibits an impressively low detection limit for the detection of SNPs at approximately 10-16 M using low-cost sensor chips and devices. By determination of the ratio of mutant DNA to normal DNA in cell-free genomic DNA from blood samples, this method is promising for diagnosing β-thalassemia in fetuses without invasive testing techniques.

Tuning assembled DNA nanoarchitecture into disassembled state to enhance accurate and ultrasensitive detection of KRAS G12D mutation

Cancer remains a formidable global health challenge, necessitating precise diagnostic tools. The KRAS gene, particularly the G12D mutation, holds pivotal clinical significance across various cancer subtypes. Existing DNA sensing techniques using single-stranded and double-stranded oligonucleotide probes are susceptible for enzymatic degradation, especially in complex biological samples like blood or serum. To address these challenges, we present an innovative approach that involves tuning assembled DNA nanoarchitecture into a disassembled state to accurate and ultrasensitive detection of the KRAS G12D mutation. Our method utilized a pair of multifunctional assembly hairpin probes to construct a DNA wire with both of a fluorophore labeled probe and a quencher labeled probe attached. Upon binding with the target DNA, the DNA wire undergoes gradual disassembly, separating all fluorophores from quenchers to significantly amplify fluorescence signals. This approach offers exceptional sensitivity and reaction kinetics, capable of detecting the KRAS G12D at femtomolar levels, and demonstrates outstanding specificity in discriminating single-base mutations. Furthermore, the DNA nanoarchitecture with well-demonstrated ability to nuclease resistance makes our method exhibits robustness in detecting KRAS G12D mutation in human serum samples. This innovative approach represents an advancement in cancer diagnostics, providing a promising tool for early disease diagnosis and biomedical study.

Label-Free Mode Based on Ferrocene/PEDOT:PSS-PPy for Molecularly Imprinted Electrochemically Ultrasensitive Detection of Amino Acids.

Generally, molecularly imprinted (MIP) electrochemical sensors for amino acids operate in a "label-like" mode. That is, after an amino acid is specifically recognized by an imprinted cavity at the sensing interface, the amino acid itself provides the sensing signal for quantitative detection. However, poorly electroactive amino acids impede electron transfer at the sensing interface and require high potentials to drive the reaction; thus, more interfering reactions tend to be triggered in practical applications, causing enhanced background noise in the detection. To address these issues, a "label-free" mode of the MIP sensor based on the ferrocene (Fc)/PEDOT:PSS-polypyrrole (PPy) composite was designed for the first time. The Fc/PEDOT:PSS-PPy is drop coated on the electrode surface as a substrate, and MIP polymers with specific recognition ability are immobilized on the substrate via electrostatic adsorption. As a proof of concept, l-tyrosine (l-Tyr) was selected as a model analyte and the "label-free" mode MIP/Fc/PEDOT:PSS-PPy sensor was constructed. The limit of detection (LOD) and linearity range of the MIP/Fc/PEDOT:PSS-PPy sensor were 2.31 × 10-11 M and from 100 pM to 5 mM, respectively. Compared with the label-like mode, the LOD was three orders of magnitude lower, the linear range was increased by three orders of magnitude, and the sensitivity was improved by more than four times. This work provides a universal and effective concept for MIP electrochemical sensing of amino acids.

Versatile and efficient fabrication of signal “turn‐on” lateral flow assay for ultrasensitive naked eye detection of small molecules based on self‐assembled fluorescent gold nanoclusters‐antigen aggregates

Versatile and efficient fabrication of signal “turn‐on” lateral flow assay for ultrasensitive naked eye detection of small molecules based on self‐assembled fluorescent gold nanoclusters‐antigen aggregates

Dual-mode optical biocompatible-sensor enabled by enzyme-like activity of UiO-66 (Zr) for ultra-sensitive ROS detection in vitro

The simple, effective and highly sensitive detection of hydrogen peroxide (H2O2), which belongs to the reactive oxygen species (ROS), at low concentrations plays an indispensable role in the field of environmental protection, biological research and safety. In this study, a dual-mode optical biosensor, UiO-66@OPD, was developed based on the inherent peroxidase mimicking activity of UiO-66 (Zr) and the optical reaction of ortho-phenylenediamine (OPD) by extending the π-system through oxidative coupling, prototropism and elimination to form OPDox, thereby exhibiting strong orangish absorbance and greenish fluorescence. The catalase-mimicking activity of UiO-66 (Zr) was demonstrated by the catalytic oxidation of methylene blue in the presence of H2O2. Moreover, the Michaelis-Menten kinetic model confirmed the intrinsic peroxidase-like activity of UiO-66@OPD as a modified MOFzyme. The synthesized UiO-66 (Zr) facilitated the oxidation of OPD to OPDox by degrading H2O2 to the hydroxyl radicals. During the oxidation process, the absorption peak at 415 nm and the fluorescence peak at 565 nm of the synthesized probe were significantly enhanced by increasing the H2O2 concentration. Moreover, a colorimetric and fluorometric ultrasensitive sensor shows a good linear relationship between the intensity enhancement and H2O2 concentration in the range of 0–600 nM for absorption and fluorescence spectra with R2 = 0.9772, and R2 = 0.9948, respectively. To demonstrate the biological performance and biocompatibility of UiO-66@OPD as a biosensor, MTT evaluation was performed for the three cell lines MCF-10 A, HEK293 and A549, indicating high biocompatibility and good cell viability for biological applications. Ultimately, this convenient, environmentally friendly, biocompatible and cost-effective catalase-mimicking-based sensor system will open a new perspective for the development of portable kite-based biosensors In vitro.

(Co, S)-Codoped BiOBr as a novel signal probe coupled with LAMP (H+)-pH responsive photoelectrochemistry biosensor for ultrasensitive detection of microRNA

Herein, Co/S-BiOBr as a novel photoelectric material with excellent photoelectric property was prepared by cation–anion co-doping method with LAMP (H+)-pH responsive as signal amplification to construct a photochemical (PEC) biosensor for ultrasensitive detection of microRNA-221 (miRNA-221) related to triple-negative breast cancer (TNBC). Notably, not only the lattice distortion caused by S doping could elevate the photo-generated electron-hole separation efficiency, but also the tune of electronic structure by Co doping promoted electron transfer and decreased recombination rate, thus significantly improving the photoelectric performance. Furthermore, in the aid of Loop-mediated isothermal amplification (LAMP), a few miRNA-221 could be converted to abundant LAMP(H+) for changing the configuration of the Fc-DNA probe, which made the ferrocene (Fc) approach to electrode via pH-responsive for hindering the transmission of electrons, resulting in a significantly reduced photocurrent signal to enhance the sensitivity of the biosensor. The proposed PEC biosensor performed a wide range from 1 fM to 10 nM with a detection limit of 0.24 fM. This work prepared a novel material with efficient photoelectric efficiency for sensitive detection of biomolecules and exhibited enormous potential in disease diagnosis.

Ultra-sensitive molecular residual disease detection through whole genome sequencing with single-read error correction

While whole genome sequencing (WGS) of cell-free DNA (cfDNA) holds enormous promise for detection of molecular residual disease (MRD), its performance is limited by WGS error rate. Here we introduce AccuScan, an efficient cfDNA WGS technology that enables genome-wide error correction at single read-level, achieving an error rate of 4.2 × 10−7, which is about two orders of magnitude lower than a read-centric de-noising method. The application of AccuScan to MRD demonstrated analytical sensitivity down to 10−6 circulating variant allele frequency at 99% sample-level specificity. AccuScan showed 90% landmark sensitivity (within 6 weeks after surgery) and 100% specificity for predicting relapse in colorectal cancer. It also showed 67% sensitivity and 100% specificity in esophageal cancer using samples collected within one week after surgery. When AccuScan was applied to monitor immunotherapy in melanoma patients, the circulating tumor DNA (ctDNA) levels and dynamic profiles were consistent with clinical outcomes. Overall, AccuScan provides a highly accurate WGS solution for MRD detection, empowering ctDNA detection at parts per million range without requiring high sample input or personalized reagents.

From Lab to Home: Ultrasensitive Rapid Detection of SARS-CoV-2 with a Cascade CRISPR/Cas13a-Cas12a System Based Lateral Flow Assay.

Currently, CRISPR/Cas-based molecular diagnostic techniques usually rely on the introduction of nucleic acid amplification to improve their sensitivity, which is usually more time-consuming, susceptible to aerosol contamination, and therefore not suitable for at-home molecular testing. In this research, we developed an advanced CRISPR/Cas13a-Cas12a-based lateral flow assay that facilitated the ultrasensitive and rapid detection of SARS-CoV-2 RNA directly from samples, without the need for nucleic acid amplification. This method was called CRISPR LFA enabling at-home RNA testing (CLEAR). CLEAR used a novel cascade mechanism with specially designed probes that fold into hairpin structures, enabling visual detection of SARS-CoV-2 sequences down to 1 aM sensitivity levels. More importantly, CLEAR had a positive coincidence rate of 100% and a negative coincidence rate of 100% for clinical nasopharyngeal swabs from 16 patients. CLEAR was particularly suitable for at-home molecular testing, providing a low-cost, user-friendly solution that can efficiently distinguish between different SARS-CoV-2 variants. CLEAR overcame the common limitations of high sensitivity and potential contamination associated with traditional PCR-based systems, making it a promising tool for widespread public health application, especially in environments with limited access to laboratory resources.

Label-free SELEX of aptamers for ultra-sensitive electrochemical aptasensor detection of amanitin in wild mushrooms

BackgroundMushroom poisoning poses a significant global health concern, with high morbidity and mortality rates. The primary lethal toxins responsible for this condition are alpha-amanitin (ɑ-AMA) and beta-amanitin (β-AMA). As a promising bio-recognition molecules in biosensors, aptamers, have been broadly used in the field of food detection. However, the current SELEX-based methods for screening aptamers for structurally similar small molecules were limited by the labelling or salt ion induction. In this study, we aimed to develop a novel label-free SELEX strategy for the screening of aptamers with high affinity and constructed new aptasensors for the detection of ɑ-AMA and β-AMA. ResultsA novel label-free SELEX strategy based on the positively charged gold nanoparticles (AuNPs) was proposed to simultaneous screening of aptamers for ɑ-AMA and β-AMA. Only 18 rounds of SELEX were required to obtain new aptamers. The candidate aptamers were analyzed by colloidal gold assay, and the sequences of ɑ-30 and β-37 displayed great affinity with Kd values of 22.26 nM and 23.32 nM, respectively, without interference from botanical toxins. Notably, the truncated aptamers ɑ-30-2 (50 bp) and β-37-2 (57 bp) exhibited higher affinity than their original counterpart (79 bp). Subsequently, the selected aptamers were utilized to construct recognition probes for electrochemical aptasensors based on hairpin cyclic cleavage of substrates by Cu2+ dependent DNAzyme and Exo I-triggered recycling cascades. The detection platform showed excellent analytical performance with limits of detection as low as 4.57 pg/mL (ɑ-AMA) and 8.49 pg/mL (β-AMA). Moreover, the aptasensors exhibited superior performance in mushroom and urine samples. SignificanceThis work developed a simple and efficient label-free SELEX method for screening new aptamers for ɑ-AMA and β-AMA, which employed the positively charged AuNPs as the screening medium, without the need for chemical labelling of libraries or induction of salt ions. Furthermore, two novel electrochemical aptasensors were developed based on our newly obtained aptamers, which offer the new biosensing tool for ultrasensitive detection of the AMA poisoning, showing great potential in practical applications.