SERS aptasensor for simultaneous detection of ochratoxin A and zearalenone utilizing a rigid enhanced substrate (ITO/AuNPs/GO) combined with Au@AgNPs.

The contamination of mycotoxins poses a serious threat to global food security, hence the urgent need for simultaneous detection of multiple mycotoxins. Herein, two SERS nanoprobes were synthesized by embedded SERS tags (4-mercaptopyridine, 4MPy; 4-mercaptobenzonitrile, TBN) into the Au and Ag core-shell structure, and each was coupled with the aptamers specific to ochratoxin A (OTA) and zearalenone (ZEN). Meanwhile, a rigid enhanced substrate Indium tin oxide glass/AuNPs/Graphene oxide (ITO/AuNPs/GO) was combined with aptamer functionalized Au@AgNPs via π-π stacking interactions between the aptamer and GO to construct a surface-enhanced Raman spectroscopy (SERS) aptasensor, thereby inducing a SERS enhancement effect for the effective and swift simultaneous detection of both OTA and ZEN. The presence of OTA and ZEN caused signal probes dissociation, resulting in an inverse correlation between Raman signal intensity (1005 cm-1 and 2227 cm-1) and the concentrations of OTA and ZEN, respectively. The SERS aptasensor exhibited wide linear detection ranges of 0.001-20 ng/mL for OTA and 0.1-100 ng/mL for ZEN, with low detection limits (LOD) of 0.94 pg/mL for OTA and 59 pg/mL for ZEN. Furthermore, the developed SERS aptasensor demonstrated feasible applicability in the detection of OTA and ZEN in maize, showcasing its substantial potential for practical implementation.

Synthesis of SnC@Au@Apta by electrospinning as an electrochemical sensor for detection of tetracycline.

A highly efficient electrochemical aptamer sensor for the detection of tetracycline (TC) was prepared by using SnC@Au@Apta. Metal tin has good electrochemical activity and high conductivity. It is often used as an electrochemical sensing material. The nanofibers prepared by electrospinning machine make the metal distribution more uniform, not easy to agglomerate, and have a certain porosity, which can improve the sensitivity of sensor detection. Carbonization further enhances conductivity. The gold nanoparticles (AuNPs) on the surface of SnC nanomaterials improve the electrochemical detection performance, and also act as the binding site of the TC aptamer, which is stably combined with the thiol group at the end of the TC aptamer. The TC aptamer specifically binds to TC to detect TC in the sample. The electrochemical performance of SnC@Au@Apta was evaluated by cyclic voltammetry (CV) and differential pulse voltammetry (DPV). Under the optimal conditions, the detection range of SnC@Au@Apta is wide (0.001-100 µM), the detection limit is low (0.83 nM), and it has excellent selectivity, stability and reproducibility. In addition, SnC@Au@Apta can be used to detect TC in milk samples.

An easy-operation aptasensor for simultaneous detection of multiple tumor-associated exosomal proteins based on multicolor fluorescent DNA nanoassemblies.

As a promising liquid biopsy biomarker, exosomes have demonstrated great potential and advantages in the noninvasive tumor diagnosis. However, an accurate and sensitive method for tumors-associated exosomes detection is scarce. Herein, we presented an easy-operation aptasensor which simultaneously detect multiple exosomal proteins by using multicolor fluorescent DNA nanoassemblies (FDNs) and CD63 aptamer-modified magnetic beads (MNPs-AptCD63). In this system, the FDNs were firstly constructed by encapsulating different quantum dots (QDs) into rolling circle amplification (RCA) products that contained different aptamer sequences. Thus, the FDNs could selectively recognize the different exosomal proteins captured by the MNPs-AptCD63, and achieve the multiplex and sensitive detection according to the fluorescence of QDs. Benefiting from the signal amplification capacity and high selectivity of FDNs, this aptasensor not only could detect exosomes as low as 650 particles/µL, but also showed accurate analysis in clinical samples. In addition, we can also achieve point-of-care testing (POCT) due to the simple analysis steps and naked-eye observable fluorescence of QDs under the ultraviolet irradiation. We believe that our aptasensor could provide a promising platform for exosomes-based personalized diagnosis and precise monitoring of human health.

Novel electrochemiluminescence platform utilizing AuNPs@Uio-66-NH2 bridged luminescent substrates and aptamers for the detection of pesticide residues in Chinese herbal medicines.

A large number of Chinese herbal medicines (CHMs) are included in daily recipes, but their pesticide residues have aroused more and more concerns. In this paper, an electrochemiluminescence aptasensor was constructed for the trace detection of acetamiprid (ACE) in Angelica sinensis and Lycium barbarum. Possessing a large specific surface area, UiO-66 was modified with amino groups to improve biocompatibility, and the addition of AuNPs allowed UiO-66-NH2 to catalyze the formation of excited states of luminescent molecules (TPrA⁎; Ru(bpy)32+⁎), and AuNPs@UiO-66-NH2 was used to bridge the aptamer (Au-S) and luminescent substrate (peptide bond). The conventional luminescent reagent Ru(bpy)32+ was doped with multi-walled carbon nanotubes (MWCNTs) to obtain a more powerful and stable light signal. After optimizing the experimental parameters, the aptasensor could give results in 10 min with a detection range from 1×10-2-1×104 nM and a lower limit of detection (LOD) of 0.8 pM. The LOD of the study was at least one order of magnitude lower than that of the fluorescence detection method. Furthermore, the accuracy of the aptasensor was validated for spiked recovery experiments.

Plasmon-enhanced fluorescence and electrochemical aptasensor for SARS-CoV-2 Spike protein detection.

In this work, we combined plasmon-enhanced fluorescence and electrochemical (PEF-EC) transduction mechanisms to realize a highly sensitive dual-transducer aptasensor. To implement two traducers in one biosensor, a novel large-scale nanoimprint lithography process was introduced to fabricate gold nanopit arrays (AuNpA) with unique fringe structures. Light transmitting through the AuNpA samples exhibited a surface plasmon polariton peak overlapping with the excitation peak of the C7 aptamer-associated fluorophore methylene blue (MB). We observed a five and seven-times higher average fluorescence intensity over the AuNpA and fringe structure, respectively, in comparison to a plane Au film. Furthermore, the MB fluorophore was simultaneously utilized as a redox probe for electrochemical investigations and is described here as a dual transduction label for the first time. The novel dual transducer system was deployed for the detection of SARS-CoV-2 Spike protein via a C7 aptamer in combination with a strand displacement protocol. The PEF transducer exhibited a detection range from 1 fg/mL to 10 ng/mL with a detection limit of 0.07 fg/mL, while the EC traducer showed an extended dynamic range from 1 fg/mL to 100 ng/mL with a detection limit of 0.15 fg/mL. This work provides insights into an easy-to-perform, large-scale fabrication process for nanostructures enabling plasmon-enhanced fluorescence, and the development of an advanced but universal aptasensor platform.

A ratiometric fluorescent biosensing platform based on CDs and AuNCs@CGO for patulin detection.

BACKGROUND: Patulin (PAT) is a mycotoxin, usually found in fruit and their products, that can potentially be harmful to human health. In order to achieve rapid detection of mycotoxins and ensure the safety of food. This study reported a novel ratiometric fluorescent aptasensor for PAT detection. In this study, we used aptamer as the recognition element, Hybrid double stranded modified with fluorescent substances as the fluorescent donor, and AuNCs@CGO as the fluorescent acceptor. After the addition of PAT, the ratiometric fluorescence "turn on" response was exhibited. RESULTS: The AuNCs@CGO are obtained by amide reaction between BSA-AuNCs and carboxylated graphene oxide (CGO). The prepared AuNCs@CGO can shorten the time of FRET effect and exhibit highly efficient quenching ability by adsorption effects (π-π stacking and electrostatic gravity) on the Aptamer-modified CDs. When the target PAT bound specifically to the CDs-apt, the fluorescence of the CDs-apt would recover, while the fluorescence of ROX modified cDNA remained unchanged. This ratiometric fluorescence response improved the accuracy of PAT detection. In addition, the proposed had good linearity for PAT in the range of 0.1-50 ng/mL with a limit of detection 0.16 ng/mL. The recovery of standard addition in grapes were 95.9%-105.4 %. SIGNIFICANCE: An effective fluorescent detection method for PAT was constructed based on aptamer and nanomaterials. This new fluorescent biosensor has the characteristics of simple synthesis, easy operation, high sensitivity, strong selectivity and a low LOD, which may be a promising idea and platform for the detection of food safety hazard factors.

An ultrasensitive free-of-electronic sacrificial agent photoelectrochemical aptasensor for the detection of dibutyl phthalate based on Z-scheme p-n Bi-doped BiOI/Bi2S3 heterojunction.

Dibutyl phthalate (DBP), a common and outstanding plasticizer, exhibits estrogenic, mutagenic, carcinogenic, and teratogenic properties. It is easily liberated from plastic materials and pollutes aquatic ecosystems, endangering human health. Therefore, highly sensitive and selective DBP detection methods are necessary. In this work, a free-of-electronic sacrificial agent photoelectrochemical (PEC) aptasensor for DBP detection was constructed using a novel Z-scheme Bi-doped BiOI/Bi2S3 (Bi-BIS) p-n heterojunction. The Bi-BIS composites had higher visible-light absorption, charge transfer, and separation efficiency. This is attributed to the synergistic effect of the formation of Z-scheme p-n heterojunction between BiOI and Bi2S3, the plasma resonance effect of metallic Bi and photosensitization of Bi2S3, thus exhibiting large and stable photocurrent response in the absence of electron sacrificial agent, that was 10.4 and 6.4 times higher than that of BiOI and Bi2S3, respectively. Then, a DBP PEC aptasensor was constructed by modifying the DBP aptamer on the surface of the ITO/Bi-BIS electrode. The aptasensor demonstrated a broad linear range (2-500 pM) and a low detection limit (0.184 pM). What's more, because there is no interference from electronic sacrificial agent, the aptasensor exhibited excellent selectivity in real water samples. Therefore, the proposed PEC has considerable potential for DBP monitoring.

Multiplex electrochemical aptasensor for the simultaneous detection of linomycin and neomycin antibiotics.

The escalating use of antibiotics across diverse sectors, including human healthcare, agriculture, and livestock, has led to their pervasive presence in the environment, raising concerns about their impact on ecosystems and human health. Traditional detection methods, reliant on high-performance liquid chromatography and immuno-assays, face challenges of complexity, cross-reactivity, and limited specificity. Aptamer-based biosensors offer a promising alternative, leveraging the specificity, stability, and cost-effectiveness of aptamers. Herein, we present a novel dual-screen-printed carbon electrode (SPCE) biosensor, modified with a nanocomposite of gold nanoparticles (AuNPs) and carbon nanofibers (CNFs), for the label-free electrochemical detection of lincomycin and neomycin antibiotics. Lincomycin and neomycin, two antibiotics of environmental concern due to their widespread usage and potential ecological impact, were simultaneously detected using square wave voltammetry. The aptasensors showed high sensitivity with detection limits of 0.02 pg/mL and 0.035 pg/mL for lincomycin and neomycin, respectively. The developed biosensor exhibited high selectivity and reproducibility in detecting both antibiotics. This multiplex biosensing platform offers a promising strategy for efficient and cost-effective monitoring of antibiotic residues in environmental samples, addressing the critical need for robust detection methods in environmental monitoring and public health surveillance.

Near-infrared-driven dual-photoelectrode photoelectrochemical sensing for fumonisin B1: Integrating a photon up-conversion bio-photocathode with an enhanced light-capturing photoanode.

Fumonisin B1 (FB1), the most prevalent and highly toxic mycotoxin within the fumonisins family, poses threats to humans, especially in children and infants, even at trace levels. Therefore, it is essential to design an easy and sensitive detection strategy. Herein, a brand-new dual-photoelectrode photoelectrochemical (PEC) sensing platform for FB1 detection under near-infrared irradiation was unveiled. This platform integrated a photon up-conversion bio-photocathode substrate (UCNPs/Au/CuInS2, UCNPs: NaYF4: Yb3+, Er3+, Nd3+) and used a SnO2/SnS2@Bi/Bi2S3 heterojunction photoanode to greatly enhance light capture. Additionally, ZnO coated with polydopamine (ZnO@PDA) was utilized as a signal inhibitor. The restoration of photocurrent occurred due to the strong binding affinity between FB1 and its aptamer (FB1-Apt), facilitating the dissociation of FB1-Apt/ZnO@PDA from the photoelectrode. The PEC sensing performance and the electron transfer process were thoroughly examined. The developed "signal-restoration" PEC aptasensor exhibited a wider dynamic linear range from 1.0 × 10-3 to 1.0 × 102 ng/mL, with a lower limit of detection (0.13 pg/mL). It has demonstrated excellent practical detection performance in unspiked real samples, such as corn paste, with the FB1 enzyme-linked immunosorbent assay (ELISA) Kit serving as a reference, indicating its potential for routine analysis of other mycotoxins. Thus, this research establishes a feasible dual-photoelectrode PEC framework for the effective detection of mycotoxins and other hazardous substances.

Double detection of mycotoxins based on aptamer induced Fe3O4@TiO2@Ag Core - Shell nanoparticles "turn on" fluorescence resonance energy transfer.

Multiple and sensitive mycotoxin detection is an essential early-warning mechanism for safeguarding human health, and preserving the environment. We synthesized a turn-on Fluorescence Resonance Energy Transfer (FRET) aptamer sensor based on the unique fluorescence quenching and substrate recognition characteristics of Ag NTs (energy receptors) and aptamer modified Fe3O4@TiO2 NP (energy donor) to detect multiple toxins using a single diagnostic approach. The addition of aflatoxin B1 (AFB1) and ochratoxin A (OTA) resulted in a change in fluorescence intensity at 510 and 650 nm, which can be employed for simultaneous recognition with detection limits of 0.94 ng·mL-1 (R2 = 0.997) and 0.54 ng·mL-1 (R2 = 0.995). The aptasensors have been successfully applied for the determination of AFB1 and OTA in grain and oil samples with high recovery rates. The approach provides novel possibilities for the development of sensitive and selective aptasensors with potential applications in aptamer-recognized multifunctional biosensing.

An autocatalytic hybridization circuit-based FRET aptasensor for detection of low-abundant sulfameter in human serum.

Exposure to antibiotics is considered a potential risk factor for human health. Yet, the extensive and cost-effective detection of low-abundant antibiotics in complex matrices remains a significant challenge. Herein, an aptamer and an autocatalytic hybridization circuit (AHC) were used to fabricate a fluorescence resonance energy transfer (FRET) platform to detect sulfameter (SME) in human serum. The AHC system comprised two mutually motivated hybridization chain reactions (HCR) modules, ultimately producing long-branched DNA copolymeric nanowires. This mutually reciprocal activation of two HCR modules enables continuous signal amplification, providing the AHC system with wide linear range and high sensitivity for the SME detection. Compared to the HCR-based aptasensor, the AHC-based aptasensor exhibited a wider linear range and improved sensitivity (3.3 times greater). Under optimal conditions, the fluorescent AHC-based aptasensor demonstrated a linear range (R2 was 0.996) from 0.5 to 2000 nM, with a low detection limit of 0.301 nM (S/N = 3). The fluorescent aptasensor was also validated by SME-spiked human serum samples, showing average recoveries ranging from 96.40 % to 109.30 %, with a relative standard deviation below 10.45 %. Furthermore, when tested on six human serum samples, the aptasensor results were consistent with those obtained from the commercial ELISA method. These findings demonstrate that the proposed aptasensor provides a promising approach for the practical monitoring of low-abundant SME in human serum.

Construction of a In2O3/ultrathin g-C3N4 S-scheme heterojunction for sensitive photoelectrochemical aptasensing of diazinon.

A single semiconductor-based photoelectrochemical (PEC) aptasensor usually faces a challenge of low sensitivity due to poor solar energy utilization and a high photogenerated carrier recombination rate. Herein, an ultra-thin carbon nitride nanosheet-coated In2O3 (In2O3/CNS) S-type heterojunction-based PEC aptasensor has been established to achieve highly sensitive detection of diazinon (DZN) pesticide in water environment. Construction of S-type heterojunction induces a band shift and an electric field effect, enhancing light utilization and accelerating directional transmission of carriers, leading to outstanding PEC performance. The creation of internal electric field at interface ensures stable carrier transport. Additionally, ultrathin CNS structure can effectively shorten the transport path of carriers. The close coating of In2O3 and CNS promotes the transfer of charge. The synergistic effects amplify the sensor's response, ultimately enabling the effective detection of DZN residue over a wide detection range (0.98 âˆ¼ 980.0 pg mL-1), a low detection limit (0.33 pg mL-1, S/N = 3) and excellent accuracy in practical application (RSD < 5 %). This work provides a reference for the construction of a new S-type heterojunction-based PEC sensor.

A dual-mode aptasensor based on rolling circle amplification enriched G-quadruplex for highly sensitive IFN-γ detection.

BACKGROUND: Aptasensors have been extensively utilized in target detection due to their advantages of high sensitivity and fast response. However, the reliability of the detection results of the single-mode aptasensor cannot be verified in time. Developing efficient detection methods with cross-validation capability is beneficial to achieving highly reliable detection. This study aims to design a colorimetric and fluorescent dual-mode aptasensor by skillfully engineering G-quadruplex assembly and rolling circle amplification for highly reliable IFN-γ detection. RESULTS: The complexes of anti-IFN-γ aptamers and complement sequences (cDNA) were modified on the magnetic beads. In the presence of IFN-γ, the preferential combination of aptamers with IFN-γ resulted in the release of cDNAs. The cDNAs were collected by magnetic separation and then used as primers to trigger rolling circle amplification reaction to generate enriched G-quadruplexes. The G-quadruplex could be utilized to combine with hemin to catalyze the oxidation of 3,3',5,5'-tetramethylbenzidine for colormetric mode or to couple with the fluorogenic dye Thioflavin T for fluorescent mode. The developed dual-mode aptasensor displayed a linear range of 1-10000 pM with a detection limit of 0.406 pM for the colormetric mode and a range of 0.1-10000 pM with a detection limit of 0.037 pM for the fluorescent mode. Further, the designed aptasensor was applied to IFN-γ detection in serum samples and achieved satisfactory recoveries. SIGNIFICANCE: This innovative dual-mode detection strategy skillfully leverages the effective target-binding ability of aptamer, dual-function of the G-quadruplex and the signal amplifying ability of rolling circle amplification. This approach not only provides a reliable testing tool for the detection of IFN-γ, but also promotes the development of multimode sensing platforms.

A HOF-101@AgNPs-based dual-signal mode aptasensor for electrochemiluminescence and fluorescence detection of E. coli O157:H7.

Escherichia coli O157:H7 (E. coli O157:H7) emerges as a foodborne pathogen, emphasizing the imperative for creating precise detection tools. An ultrasensitive electrochemiluminescence/fluorescence (ECL/FL) dual-signal mode aptasensor was constructed for the detection of E. coli O157:H7. Ultrafine silver nanoparticles were loaded on the surface of hydrogen-bonded organic skeleton materials by in-situ photoreduction method, and then combined with aptamers that can identify specific targets to prepare HOF-101@AgNPs@Apt, greatly simplifying the ECL/FL dual-signal mode probe fabrication process and improving sensing reliability. HOF-101@AgNPs@Apt had both strong ECL luminescence and fluorescence, resulting in a high detection sensitivity. The low limit of detection (LOD) for ECL and FL were 0.48 CFU mL-1 and 2.39 CFU mL-1, respectively. Moreover, the proposed dual-signal mode aptasensor has been successfully applied to determine E. coli O157:H7 levels in tap water and milk with superior accuracy and high antiinterference capability, providing a promising method for food safety monitoring.

Laser-induced graphene-based aptasensor for the selective detection of Escherichia coli in urine samples.

A Laser-Induced Graphene-based (LIG) electrode covalently functionalized with an aptamer (P12-55) was used to develop an aptasensor detecting Escherichia coli in urine samples. Recent strides in material science have spotlighted LIG for exceptional attributes like robust mechanical resistance, superior conductivity, extensive surface area, and facile synthesis/patterning on various polymeric substrates. Variations in the aptasensor charge transfer resistance upon interaction with bacterial cells were evaluated by electrochemical impedance spectroscopy. Tests in phosphate buffer saline solution showed the aptasensor linear response for E. coli between 100 and 103 CFU/mL. The sensor proved to be selective for E. coli as a negligible response was observed in the presence of Staphylococcus aureus or Pseudomonas aeruginosa. Finally, the sensor was calibrated in urine samples spiked with a known concentration of E. coli cells. Its characteristics make the aptasensor viable for low-cost and portable devices identifying pathogenic microorganisms at the point-of-need. Due to the short response time (about 30 min), we believe that these sensing devices may significantly improve control and management of urinary tract infections.

Enhancing resolution in DNA staining dye-based label-free visual fluorescence aptasensor: Strategy for eliminating non-specific binding-induced signal interference.

By optimizing the quenching capabilities of diverse two-dimensional (2D) nanomaterials such as graphene oxide (GO), Ti3C2 MXene, and MoS2, we have pioneered a label-free fluorescence aptasensor with near-zero background signal, enabling highly sensitive detection of aflatoxin B1 (AFB1). This aptasensor was equipped with a newly synthesized dicationic fluorophore, VLM, which exhibited binding-induced turn-on fluorescence properties. Among the evaluated 2D nanosheets, MoS2 nanosheets were found to exhibit exceptional quenching efficiency for the background emission of the cDNA/VLM complex (cDNA was the complementary DNA of the aptamer), further enhancing the overall performance of our aptasensor. Upon exposure to AFB1, the aptamers underwent conformational switching and target binding, leading to the formation of aptamer/AFB1 complex. Additionally, the aptamers bound complementarily to cDNA, creating aptamer-cDNA duplexes that interacted with VLM, resulting in a robust fluorescence signal. Despite the presence of a weakly fluorescent cDNA/VLM background, this fluorescence could be effectively quenched by the addition of MoS2 nanosheets. Consequently, the label-free fluorescence aptasensor exhibited excellent linearity with AFB1 concentration within 2-3000 ng mL-1, achieving a limit of detection (LOD) of 0.006 ng mL-1. Remarkably, the visual fluorescence captured by a smartphone camera can be processed using extracted grayscale values, consistently revealing a linear relationship with the AFB1 concentration within 2-3000 ng mL-1, with a LOD of 0.197 ng mL-1. This aptasensor demonstrated exceptional sensitivity and a remarkably rapid sample-to-answer detection time of 74 min, showcasing its immense potential as a straightforward, sensitive, and visually intuitive method for rapid AFB1 detection with enhanced resolution.

Development of a label-free electrochemical aptasensor for Rift Valley fever virus detection.

In this research, we describe the first aptasensor for the detection of the Rift Valley Fever virus (RVFV). The process involved the selection of aptamers through the systematic evolution of ligands by the exponential enrichment (SELEX) technique. After 12 rounds of selection, 6 aptamers were selected and the corresponding binding affinities were assessed using fluorescence binding assays, revealing dissociation constants ranging from 15.45 to 40.98 nM. Notably, among the aptamers, RV2 and RV3 exhibited the highest binding affinities toward RVFV, with dissociation constants of 15.45 and 18.62 nM, respectively. Thiol-modified aptamers were subsequently immobilized onto screen-printed gold electrodes, facilitating the label-free detection of RVFV through square wave voltammetry. The voltammetric aptasensor demonstrated an excellent sensitivity, with a detection limit of 0.015 ng/mL. In addition, cross-reactivity assessments were conducted, where negligible response was obtained when the aptasensor was exposed to non-specific proteins.

Brevetoxin Aptamer Selection and Biolayer Interferometry Biosensor Application.

Brevetoxins (PbTxs) are very potent marine neurotoxins that can cause an illness clinically described as neurologic shellfish poisoning (NSP). These toxins are cyclic polyether in chemistry and have increased their geographical distribution in the past 2 decades. However, the ethical problems as well as technical difficulties associated with currently employed analysis methods for marine toxins have spurred the quest for suitable alternatives to be applied in a regulatory monitoring regime. In this work, we reported the first instance of concurrent aptamer selection of Brevetoxin-1 (PbTx-1) and Brevetoxin-2 (PbTx-2) and constructed a biolayer interferometry (BLI) biosensor utilizing PbTx-1 aptamer as a specific recognition element. Through an in vitro selection process, we have, for the first time, successfully selected DNA aptamers with high affinity and specificity to PbTx-1 and PbTx-2 from a vast pool of random sequences. Among the selected aptamers, aptamer A5 exhibited the strongest binding affinity to PbTx-1, with an equilibrium dissociation constant (KD) of 2.56 µM. Subsequently, we optimized aptamer A5 by truncation to obtain the core sequence (A5-S3). Further refinement was achieved through mutations based on the predictions of a QGRS mapper, resulting in aptamer A5-S3G, which showed a significant increase in the KD value by approximately 100-fold. Utilizing aptamer A5-S3G, we fabricated a label-free, real-time optical BLI aptasensor for the detection of PbTx-1. This aptasensor displayed a broad detection range from 100 nM to 4000 nM PbTx-1, with a linear range between 100 nM and 2000 nM, and a limit of detection (LOD) as low as 4.5 nM. Importantly, the aptasensor showed no cross-reactivity to PbTx-2 or other marine toxins, indicating a high level of specificity for PbTx-1. Moreover, the aptasensor exhibited excellent reproducibility and stability when applied for the detection of PbTx-1 in spiked shellfish samples. We strongly believe that this innovative aptasensor offers a promising alternative to traditional immunological methods for the specific and reliable detection of PbTx-1.

An ultrasensitive liquid crystal aptasensing chip assisted by three-way junction DNA pockets for acrylamide detection in food samples.

Acrylamide, an unsaturated amide found in heat-processed foods, poses serious risks to human health due to its neurotoxicity, carcinogenicity, and genotoxicity. This highlights the importance of quantitative determination of acrylamide in foods and the environments to ensure public health safety. Therefore, there is an urgent need for simple, rapid, and highly sensitive methods to accurately quantify acrylamide. In the present study, a user-friendly aptasensor was designed to quantify ultra-low levels of acrylamide in nuts for the first time. This innovative approach utilizes chemical engineering of a glass slide as a portable sensing platform, which incorporates liquid crystal (LC) molecules and a three-way junction (TWJ) DNA pocket. The immobilized TWJ pocket can disrupt the vertical alignment of LCs, turning the dark polarized background of the aptasensor to a colorful state. The binding of the specific aptamer to acrylamide disrupts the TWJ structure, enabling the LCs to return to their homotropic alignment. This structural change restores the dark polarized view of the sensing platform. The TWJ-engineered LC aptasensor effectively detects ultra-low concentrations of acrylamide in the range of 0.0005 to 50 fmol/L, with a detection limit of 0.106 amol/L. The aptasensor was successfully applied to real roasted nut samples, including peanut, almond, pistachio, and hazelnut, achieving recovery values ranging from 96.84 % to 99.61 %. With its simplicity, portability, ease of use, and cost-effectiveness, this aptasensor is a powerful sensing device for food safety monitoring.

Development of transition metal oxide platforms for aptasensing of PSA in cell cultures.

In this study, a novel aptasensor based on a transition metal oxide-modified pencil graphite electrode (PGE) was developed for the diagnosis of early-stage prostate cancer (PCa) via monitoring the prostate-specific antigen (PSA), which is the main biomarker for PCa. Single-use PGEs modified with pulsed deposited manganese oxide (MnOx) film were used to attach the amino-terminated aptamer specific to the PSA via carbodiimide chemistry. The designed aptasensor was placed in an electrochemical cell containing ferri/ferrocyanide ions as a redox probe to measure the charge transfer resistances (Rct) of the electrode surface by electrochemical impedance spectroscopy (EIS) to follow the response of each modification step. The effect of the medium pH on the ionic structure of the aptamer molecule according to its pI value and, thus, the reversing of the direction of the response (ΔRct) by the pH change was also discussed. The level of PSA secreted from PCa cells was investigated using impedimetric transduction. The specificity of the aptasensor was validated through selectivity studies against non-specific tumor markers like VEGF and different cancer cell lines including breast cancer and androgen-insensitive prostate cancer. The developed system showcases a label-free, fast, specific, and cost-effective approach for PSA detection, highlighting the importance of medium pH and the electrostatic environment on the aptamer's response. Our work emphasizes the potential for such aptasensors in clinical diagnostics and paves the way for further exploration into using transition metal oxides in biosensing applications.