Detection Of Okadaic Acid Research Articles

A smartphone-assisted ultrasensitive colorimetric aptasensor based on DNA-encoded porous MXene nanozyme for visual detection of okadaic acid

The smartphone-assisted ultrasensitive colorimetric aptasensor based on DNA-encoded porous Ti3C2 nanozyme (Apt-P-Ti3C2) was exploited for real-time detection of OA. Porous Ti3C2 (P-Ti3C2) MXene with outstanding peroxidase-like activities were crafted using microwave combustion, facilitating the efficient catalysis of chromogenic substrate oxidation by H2O2. The integration of a considerable number of unsaturated Ti center edges and residual Mn2+ within the single-layer porous Ti3C2 framework augmented the adsorption capacity of DNA aptamer, thereby yielding a heightened catalytic efficacy of P-Ti3C2 nanoparticles. The enhanced catalytic activity of P-Ti3C2 can be partially diminished through specific recognition of OA. Concurrently, a smartphone platform was integrated for signal reading based on the colorimetric sensing strategy. The smartphone-based biosensor exhibited a reliable and ultrasensitive capability for OA detection with the detection limit of 0.38 ng·mL−1. It is anticipated that the developed smartphone-based biosensing platform can provide a prospective ultrasensitive detection method for marine algal toxin in food analysis.

Integrated OPECT and Smartphone Colorimetry Dual‐Mode Detection of Okadaic Acid Based on Ce‐MOF Modified MXene@SnO2 Z‐Scheme Heterostructure

AbstractThe organic photoelectrochemical transistor (OPECT) biosensing relies solely on a singular signal readout inherently, which restrains the precision and dependability nestled within pertinent biological measurements. Herein, a high‐precision magnetic assisted OPECT and smartphone colorimetric (SCL) dual‐mode biosensing platform is first established for detecting harmful algal toxin okadaic acid (OA) by biocatalytic reaction. MXene@SnO2‐Ce‐MOF (MXSnO/Ce‐MOF) Z‐scheme heterojunctions with abundant oxygen vacancies are prepared as photoactive materials. Initially, in the presence of OA, the coupling of trigger DNA (tDNA) to magnetic beads (MBs) via anchor DNA (aDNA) is released through the interaction of the target analyte with the aptamer. Subsequently, the carried tDNA triggers HCR between the two hairpin sequences, producing long double helix chains to capture glucose oxidase (GOx). The obtained GOx supernatant catalyzes glucose to produce H2O2, which can oxidize Ce‐MOF, leading to the alteration of electrode color and a significant decrease in the overall photocurrent of MXSnO/Ce‐MOF. Crucially, the novel OPECT‐SCL biosensor exhibits excellent sensitivity and precision, boasting detection thresholds as low as 42.9 pM and 1.2 nM, respectively, and accomplishes the automated detection of OA within real samples. The proposed OPECT‐SCL dual‐signal measurement model constitutes a sensitive, portable, and precise platform for the quantification of marine toxins.

Controlled Spread of a Ag Layer from the Core to the Tip along the Branches of AuAg Nanostars for Improved SERS Detection of Okadaic Acid in Shellfish.

Plasmonic Au-Ag nanostars are excellent surface-enhanced Raman scattering (SERS) probes due to bimetallic coupling and the tip effect. However, the existing preparation methods of AuAg nanostars cannot achieve controlled growth of the Ag layer on the branches of nanostars and so cannot display their SERS to the maximum extent, thus limiting its sensitivity in biosensing. Herein, a novel strategy "PEI (polyethylenimine)-guided Ag deposition method" is proposed for synthesizing AuAg core-shell nanostars (AuAg@Ag NS) with a tunable distribution of the Ag layer from the core to the tip, which offers an avenue for investigating the correlation between SERS efficiency and the extent of spread of the Ag layer. It is found that AuAg@Ag NS with a Ag layer coated the whole branch has the strongest SERS performance because the coupling between the tips and Ag layer is maximized. Meanwhile, as a completely closed core-shell structure, AuAg@Ag NS can confine and anchor 4-ATP inside the Ag layer to avoid an unstable SERS signal. By connecting the aptamer, a reliable internal standard nanoprobe with a SERS enhancement factor (EF) up to 1.86 × 108 is prepared. Okada acid is detected through competitive adsorption of this SERS probes, and the detection limit is 36.6 pM. The results gain fundamental insights into tailoring the nanoparticle morphologies and preparation of internal standard nanoprobes and also provide a promising avenue for marine toxin detection in food safety.

MXene@MnIn2S4‐Gated Organic Photoelectrochemical Transistors with Nanozyme‐Mediated Multiple Quenching Effects for Ultrasensitive Detection of Okadaic Acid

AbstractOrganic optoelectronics have attracted widespread interdisciplinary research interest but lags far behind in the application in marine environmental detection. The organic photoelectrochemical transistor (OPECT) shows promise as a powerful tool for comprehensive monitoring and early warning of marine conditions, which can be further enhanced by the valuable signal amplification strategy of nanozyme‐mediated catalytic precipitation. Herein, OPECT technology is integrated with nanozyme‐mediated catalytic precipitation for the first time, establishing an ultrasensitive detection platform for okadaic acid (OA). Specifically, MXene@MnIn2S4 (MXMnIS) hybrid composed of Schottky‐junction is synthesized via a hydrothermal method, which can efficiently modulate the device with high current gain. Linking with a sandwich immunoassay, the Ru‐C3N4 nanozyme with peroxidase‐mimicking activity can catalyze the oxidation of 4‐chloro‐1‐naphthol (4‐CN) to form an insoluble precipitate on the electrode surface, resulting in a decrease in the photocurrent and altering the transistor response. Importantly, the proposed OPECT biosensor presented an excellent sensitivity and a low detection limit (32.5 pM), fully satisfying the fundamental requirements for the quantitative detection of intracellular and extracellular OA in the practical culture media of Prorocentrum lima at different growth stages. This OPECT platform based on the nanozyme‐mediated quenching effect is significant for effectively monitoring the safety of the marine ecological environment and food safety.

A graphene oxide-assisted protein immobilization paper-tip immunosensor with smartphone and naked eye readout for the detection of okadaic acid

BackgroundOkadaic acid (OA), as a diarrhetic shellfish poisoning, can increase the risk of acute carcinogenic or teratogenic effects for the ingestion of OA contaminated shellfish. At present, much effort has been made to graft immunoassay onto a paper substrate to make paper-based sensors for rapid and simple detection of shellfish toxin. However, the complicated washing steps and low protein fixation efficiency on the paper substrate need to be further addressed. ResultsA novel paper-tip immunosensor for detecting OA was developed combined with smartphone and naked eye readout. The trapezoid paper tip was consisted of quantitative and qualitative detection zones. To improve the OA antigen immobilization efficiency on the paper substrate, graphene oxide (GO)-assisted protein immobilization method was introduced. Meanwhile, Au nanoparticles composite probe combined with the lateral flow washing was developed to simplify the washing step. The OA antigen-immobilized zone, as the detection zone Ⅰ, was used for quantitative assay by smartphone imaging. The paper-tip front, as the detection zone Ⅱ, which could qualitatively differentiate OA pollution level within 45 min using the naked eye. The competitive immunoassay on the paper tip exhibited a wide linear range for detecting OA (0.02–50 ng∙mL−1) with low detection limit of 0.02 ng∙mL−1. The recovery of OA in spiked shellfish samples was in the range of 90.3 %–113.%. SignificanceThese results demonstrated that the proposed paper-tip immunosensor could provide a simple, low-cost and high-sensitivity test for OA detection without the need for additional large-scale equipment or expertise. We anticipate that this paper-tip immunosensor will be a flexible and versatile tool for on-site detecting the pollution of marine products.

An electrochemiluminescent dual-signal immunosensor based on a Ru(bpy)32+-doped silica nanoparticle/copper nanocluster composite for okadaic acid detection

In this study, a sensitive dual-signal electrochemiluminescence (ECL) immunosensor was developed for okadaic acid (OA) detection utilizing copper nanoclusters (CuNCs) and Ru(bpy)32+-doped silica nanoparticles (RuSiNPs). Interestingly, the CuNCs could simultaneously enhance both cathodic (−0.95 V) and anodic (+1.15 V) ECL signals of RuSiNPs, forming a dual-signal ECL sensing platform. Further, RuSiNPs@CuNCs were used as immunomarkers by covalently conjugating them with an anti-OA monoclonal antibody (mAb) to form probes. Finally, dual ECL signals of the immunosensor were fabricated and showed good linear relationships with OA concentrations in the range of 0.05–70 ng mL−1, having a median inhibitory concentration (IC50) of 1.972 ng mL−1 and a limit of detection of 0.039 ng mL−1. Moreover, the constant ratio of the cathodic and anodic ECL peaks achieved self-calibration of the detection signal and improved the reliability of the results. Finally, we successfully applied the ECL sensor to detect OA in spiked oyster samples.

A 2D carbon nitride-based electrochemical aptasensor with reverse amplification for highly sensitive detection of okadaic acid in shellfish.

Okadaic acid (OA) is one of the main virulence factors of diarrheal shellfish toxins (DSP). It is of great significance to detect OA with an accurate, specific and cost-effective technique in the fields of seafood safety and water quality control. In this work, an electrochemical aptasensor with reverse amplification was developed for the sensitive detection of OA. A two-dimensional graphite-phase nanomaterial (carbon nitride) modified with an anti-OA aptamer and thionine (Th) was immobilized onto the surface of the electrochemical electrode as the sensitive element to capture target OA molecules. ssDNA-modified carbon nitride was used as the reverse amplification element by hybridizing with non-OA linked aptamers. The preparation of the electrochemical aptasensor was well characterized by Scanning Electron Microscopy (SEM), zeta potential detection, UV-Vis absorption, Brunner-Emmet-Teller (BET) measurements, and electrochemical measurements. The quantitative assessment of OA was achieved by differential pulse voltammetry (DPV). Experimental results indicated that this aptasensor showed a concentration-dependent response to OA with a good detection performance including in terms of selectivity, repeatability, reproducibility, and stability. It exhibited 100-fold selectivity between OA and other toxins including dinophysistoxins (DTX), pectenotoxins (PTX), and yessotoxins (YTX). In addition, it showed a much wider quantification range, which is 10-13 M-10-10 M (0.080-80.50 pg mL-1). The detection limit was as low as 10-13 M (0.080 pg mL-1). The aptasensor also successfully achieved significant practicality on real shellfish samples contaminated by OA. All these results demonstrated that the reverse amplification strategy for marine toxin detection may provide a label-free and rapid detection approach for portable applications in the fields of environmental monitoring and food security.

Correction: Highly sensitive solid-state nanopore aptasensor based on target-induced strand displacement for okadaic acid detection from shellfish samples

Correction for ‘Highly sensitive solid-state nanopore aptasensor based on target-induced strand displacement for okadaic acid detection from shellfish samples’ by Mohamed Amin Elaguech et al., Sens. Diagn., 2023, 2, 1612–1622, https://doi.org/10.1039/D3SD00199G.

Antibody-free and selective detection of okadaic acid using an affinity peptide-based indirect assay.

Okadaic acid (OA) is a type of marine biotoxin produced by some species of dinoflagellates in marine environments. Consumption of shellfish contaminated with OA can cause diarrhetic shellfish poisoning (DSP) in humans with symptoms that typically include abdominal pain, diarrhea and vomiting. In this study, we developed an affinity peptide-based direct competition enzyme-linked immunosorbent assay (dc-ELISA) for the detection of OA in real samples. The OA-specific peptide was successfully identified via M13 biopanning and a series of peptides were chemically synthesized and characterized their recognition activities. The dc-ELISA system showed good sensitivity and selectivity with a half-maximal inhibitory concentration (IC50) of 148.7ng/mL and a limit of detection (LOD) of 5.41ng/mL (equivalent, 21.52ng/g). Moreover, the effectiveness of the developed dc-ELISA was validated using OA-spiked shellfish samples, and the developed dc-ELISA showed a high recovery rate. These results suggest that the affinity peptide-based dc-ELISA can be a promising tool for detecting OA in shellfish samples.

Mismatched duplexed aptamer-isothermal amplification-based nucleic acid-nanoflower for fluorescent detection of okadaic acid.

We developed a highly sensitive fluorescent assay to detect okadaic acid (OA), a prevalent aquatic toxin posing serious health risks. Our approach uses a mismatched duplexed aptamer (DA) immobilized on streptavidin-conjugated magnetic beads (SMBs) to create a DA@SMB complex. In the presence of OA, the cDNA unwinds, hybridizes with a G-rich segment pre-encoding circular template (CT), and undergoes rolling circle amplification (RCA) to produce G-quadruplexes, which are detected using the fluorescent dye thioflavine T (ThT). The method has a LOD of 3.1×10-3 ng/mL, a linear range of 0.1∼1.0×103 ng/mL, and was successfully applied to shellfish samples with spiked recoveries of 85.9% ∼ 102.2% and RSD less than 13%. Furthermore, instrumental analysis confirmed the accuracy and reliability of this rapid detection method. Overall, this work represents a significant advancement in the field of rapid aquatic toxin detection and has important implications for public health and safety.

Efficient capture and highly sensitive analysis of okadaic acid by three-dimensional covalent organic frameworks with hydroxyl surface engineering

A novel three-dimensional covalent organic framework (3D-COF) with content-tunable and active hydroxyl groups (OH) on the pore walls was developed and adopted for the high-performance capture of okadaic acid (OA) marine toxins. Using pore-surface engineering, the integration of linear building blocks (4,4′-diamino-3,3′-biphenyldiol, BD(OH)2 and benzidine, BD) with the 3D structural building block backbone (4,4′,4′',4′''-methane-tetrayltetrabenzaldehyde, TFPM) was achieved. By adjusting the ratio of BD(OH)2, functional multicomponent-COFs [OH]x-BD-TFPM COFs (X = 25%) were synthesized, which offered ideal access to convert a conventional COF into a functional platform with multiple-mode interactions of hydrophobic and hydrophilic groups for OA capture. [OH]x-BD-TFPM was characterized using SEM, XRD, FT-IR, and BET. The adsorption features and analytical performance of OA were screened and evaluated. Optimization of dispersive solid-phase extraction using [OH]25-BD-TFPM was accomplished, and the method was verified for sensitive quantitative detection of OA in clam and mussel samples. Coupled with LC-MS/MS, the resultant [OH]25-BD-TFPM COF demonstrated the ability to analyze OA, and the limit of detection for OA in shellfish was determined to be 0.005 μg/kg. A significant improvement in trace OA detection was observed compared to previously reported SPE materials without adjustable hydrophilic interactions. The recoveries of OA in the fortified clam and mussel samples were in the ranges of 93.9‒105.1% and 96.7‒110.2%, respectively. This study highlights that OH-group surface engineering in channel walls is a facile and powerful strategy for developing functional 3D-COFs with multiple interactions for high-performance target capture.

Okadaic Acid Detection through a Rapid and Sensitive Amplified Luminescent Proximity Homogeneous Assay.

Okadaic acid (OA), a marine biotoxin produced by microalgae, poses a significant threat to mariculture, seafood safety, and human health. The establishment of a novel, highly sensitive detection method for OA would have significant practical and scientific implications. Therefore, the purpose of this study was to develop an innovative approach for OA detection. A competitive amplified luminescent proximity homogeneous assay (AlphaLISA) was developed using the principle of specific antigen-antibody binding based on the energy transfer between chemiluminescent microspheres. The method was non-washable, sensitive, and rapid, which could detect 2 × 10-2-200 ng/mL of OA within 15 min, and the detection limit was 4.55 × 10-3 ng/mL. The average intra- and inter-assay coefficients of variation were 2.54% and 6.26%, respectively. Detection of the actual sample results exhibited a good correlation with high-performance liquid chromatography. In conclusion, a simple, rapid, sensitive, and accurate AlphaLISA method was established for detecting OA and is expected to significantly contribute to marine biotoxin research.

NIR-responsive photoelectrochemical sensing platform for the simultaneous determination of tetrodotoxin and okadaic acid in Nassariidae

Simultaneous detection of tetrodotoxin (TTX) and okadaic acid (OA) is important for seafood safety. In this work, a novel paper electrode-based near-infrared (NIR) light-responsive photoelectrochemical (PEC) immunosensor was constructed using Ag2S quantum dots (QDs) and NaYF4: Yb, Er upconversion nanoparticles (UCNPs) matched with BiOI for the simultaneous detection of TTX and OA in aquatic products. A low-cost, easily prepared gold nanoparticle-functionalized paper-based screen-printed electrode with six channels was designed to immobilize OA and Ab1 of TTX. Correspondingly, PEC signal immunoprobes (BiOI@UCNPs-Ab and Ab2-Ag2S QDs) with NIR-light response were introduced to construct competitive-type and sandwich-type PEC immunosensors for OA and TTX, respectively. Under optimal conditions, the linear ranges for TTX and OA were 0.001–100 and 0.001–80 ng mL−1, respectively, and the detection limits were 5 and 7 pg mL−1, respectively. The proposed sensor was successfully used for the simultaneous analysis of TTX and OA in Nassariidae samples.

Oriented-aptamer encoded magnetic nanosensor with laser-induced fluorescence for ultrasensitive test of okadaic acid

Okadaic acid (OA) biotoxin acts a well-established inhibitor of protein phosphatase even a tumor promoter of human being, arouse great attention in safety monitoring. However, the powerful and convenient nanosensing technologies for addressing the demands such as rapidity, high sensitivity, and stability in the in-field test of OA shellfish toxin is still scarce. Herein, a high-performance magnetic biometric nanosensor (MBNS) integrating oriented aptamers and ultrasensitive laser-induced fluorescence (LIF) was firstly proposed for the in-field detection of trace OA in seafoods. High-density aptamers hybridized with FAM-labeled cDNA were tethered to the surface of AuNPs on magnetic MIL-101@Fe3O4, and then finely regulated by mercaptohexyl alcohol (MCH) to be orderly assembled, as was successfully utilized to engineer an active biological nanosensor for highly specific recognition of OA. Aptamers anchored on magnetic Fe3O4@MOF@AuNPs activate a biometric microreactor of OA, in which the superior LIF properties, conformation regulation of aptamer, and the specific recognition using aptamer genes were adopted. The magnetic nanosensor with an excellent specificity and super sensitivity for OA analysis was achieved within 20 min. Moreover, the content of captured OA could facilely be recorded by measuring the fluorescence intensity, and the limit of detection (LOD) and limit of quantitation of OA (LOQ) reached 0.015 and 0.050 ng/mL respectively, which was far better than most aptamer-based biometric sensing methods. The feasibility for accurate test of trace OA toxin in the fortified shellfish samples was validated with the recovery yields of 88.2–107.5% and RSD of 0.5–7.6%, respectively. The result demonstrated that the oriented-aptamer encoded MNS had significant practical values in rapid and ultrasensitive detection of OA biotoxin and the related safety applications.

Rapid quantitative detection of okadaic acid in shellfish using lanthanide-labelled fluorescent-nanoparticle immunochromatographic test strips

Rapid quantitative detection of okadaic acid in shellfish using lanthanide-labelled fluorescent-nanoparticle immunochromatographic test strips

Triple-helix molecular switch-based light-addressable potentiometric aptasensor for the multi-channel highly sensitive label-free detection and spatiotemporal imaging of okadaic acid

Okadaic acid (OA) has attracted considerable attention in the fields of human health and public safety because it can cause severe virulence and is widely distributed. Therefore, it is of immediate significance to explore a novel method for highly sensitive, low-consumption, efficient, and convenient OA detection. In this study, a triple-helix molecular switch (THMS)-based light-addressable potentiometric aptasensor (LAPA) is proposed for the highly sensitive and selective label-free detection and spatiotemporal imaging of OA. In this method, a THMS containing an OA aptamer served as the recognition element, and a DNA microarray including a light-addressable potentiometric sensor was designed as the transducer. Meanwhile, poly (allylamine hydrochloride) was modified on the chip surface to enhance DNA absorption and signal detection. A detection platform was constructed to measure signals and scan spatiotemporal images. The realized biosensing system enabled the multichannel detection of OA in a linear range of 0.05–100 nM with low sample consumption, a detection limit of 0.03 nM, and a detection time of 27 min/channel and spatiotemporal imaging analysis with a resolution of 400 µm2/pixel and frame duration of 3.2 min/frame. Considering the technical developing of aptamer-based THMS and micro/nano biosensor, the proposed method will provide a promising approach for label-free detection and spatiotemporal analysis of OA with high sensitivity, specificity, and detection efficiency; low sample dosage; and simple operation.

A Sensitive Fluorescence Polarization Immunoassay for the Rapid Detection of Okadaic Acid in Environmental Waters

In this study, a homogeneous fluorescence polarization immunoassay (FPIA) for the detection of hazardous aquatic toxin okadaic acid (OA) contaminating environmental waters was for the first time developed. A conjugate of the analyte with a fluorophore based on a fluorescein derivative (tracer) was synthesized, and its interaction with specific anti-OA monoclonal antibodies (MAbs) was tested. A MAbs–tracer pair demonstrated highly affine immune binding (KD = 0.8 nM). Under optimal conditions, the limit of OA detection in the FPIA was 0.08 ng/mL (0.1 nM), and the working range of detectable concentrations was 0.4–72.5 ng/mL (0.5–90 nM). The developed FPIA was approbated for the determination of OA in real matrices: river water and seawater samples. No matrix effect of water was observed; therefore, no sample preparation was required before analysis. Due to this factor, the entire analytical procedure took less than 10 min. Using a compact portable fluorescence polarization analyzer enables the on-site testing of water samples. The developed analysis is very fast, easy to operate, and sensitive and can be extended to the determination of other aquatic toxins or low-molecular-weight water or food contaminants.

Single-stranded DNA binding protein coupled aptasensor with carbon-gold nanoparticle amplification for marine toxins detection assisted by a miniaturized absorbance reader

Okadaic acid (OA), one of the most widely distributed marine toxins worldwide poses a severe threat to human health. Previous sensing methods for OA detection are usually based on antigen-antibody binding mechanism. However, the drawbacks of antibodies especially the enzyme-labeled antibodies, such as the harsh storage condition and high cost, lead to significant challenges to OA detection in biological samples. To overcome these limitations, a single-stranded DNA binding protein (SSB) coupled aptasensor was developed for OA detection. SSB was incubated on the microplate as a substitute for conventional OA-protein conjugations. Carbon-gold nanoparticles were synthesized and labeled with horseradish peroxidase and thiol-modified aptamers to obtain a capture probe (CGNs@HRP-Apt) instead of the enzyme-labeled antibody for signal amplification. OA and SSB competed to bind with limited aptamers on CGNs@HRP-Apt probes followed by colorimetric assay to obtain the optical signals correlated to OA concentration. To achieve on-site detection, a miniaturized and multichannel absorbance reader (Smart-plate reader) was self-designed with full automation for OA detection. Utilizing the SSB coupled aptasensor and the Smart-plate reader, our approach enables cost-effective and on-site OA sensing with a detection range of 2.5–80 ppb and an ultra-low limit of detection of 0.68 ppb. Moreover, novel OA detection kits based on the SSB coupled aptasensor were prepared which can effectively reduce the cost by 15 times lower than that of commercial ELISA kits. Therefore, the developed platform provides a favorable and promising avenue for marine toxin detection in aquaculture and food safety.

Antibody-Free and Selective Detection of Okadaic Acid Using an Affinity Peptide-Based Indirect Assay

Antibody-Free and Selective Detection of Okadaic Acid Using an Affinity Peptide-Based Indirect Assay

Highly sensitive solid-state nanopore aptasensor based on target-induced strand displacement for okadaic acid detection from shellfish samples

Okadaic acid (OA) is a marine toxin that is frequently found in a wide variety of shellfish and can cause major health complications.