50 results about "Okadaic acid" patented technology

The invention discloses a method for detecting diarrhea shellfish toxin, which comprises the following steps: A) establishing a standard curve of depolymerization of okadaic acid (OA) induced HL-7702 hepatocyte F-actin; B) carrying out the calculation on the concentration of the OA according to the standard curve of the F-actin depolymerization of the HL-7702 hepatocyte, establishing a detection method of the hepatocyte F-actin of the diarrhea shellfish toxin to determine the possible detection limit range; C) selecting one or more of components (STX, DA and YTX) which are possibly coexist with the diarrhea shellfish to act on the HL-7702 hepatocyte, determining the specificity of the method for detecting the OA content by detecting the degree of damaging the polymerizing power of the HL-7702 hepatocyte F-actin by the above components. The detection method of the invention has the advantages: (1) taking cells as experimental subjects to avoid the using of experimental animals and conforming to the rule of 3R; (2) having high sensitivity; (3) having good specificity; (4) having good repeatability; and (5) having simple and convenient sample extraction and low matrix effects.

The present invention provides an inhibitor of NF-κB, guggulsterone and its analogs. Guggulsterone suppresses NF-κB activation induced by TNF, phorbol ester, okadaic acid, cigarette smoke, H2O2 and IL-1β, as well as constitutive NF-κB activation expressed in most tumor cells. One mechanism by which guggulsterone inhibits activation of NF-κB is through suppression of IκBα phosphorylation and IκBα degradation. NF-κB-dependent gene transcription is modulated by guggulsterone and its analogs. In particular, induction by TNF, TNFR1, TRADD, TRAF2, NIK and IKK, is modulated by guggulsterone and its analogs. In addition, guggulsterone decreased the expression of genes involved in anti-apoptosis (IAP1, XIAP, Bfl-1 / A1, bcl-2, cFLIP, survivin), proliferation (cyclin D1, c-myc) and metastasis (MMP-9, COX2 and VEGF).

The invention provides a group of single-chain DNA nucleic acid aptamers for specifically recognizing okadaic acid and belongs to the biotechnical field of food safety. A group of nucleic acid aptamers are obtained through 13 rounds of vitro screening of incubating, separating, amplifying and preparing single chains by virtue of a systematic evolution technology of ligands by exponential enrichment employing graphene oxide-assisted separation; and an aptamer OA-27 which has high affinity and high specificity to the okadaic acid is screened through affinity and specificity analysis. Through analysis, OA27-1 from which a primer zone is removed also has high affinity and specificity. The single-chain DNA nucleic acid aptamers have a wide application prospect in analysis detection of the okadaic acid in aquatic products and diagnosis and treatment of okadaic acid poisoning in clinical.

The invention discloses an electrochemical detection method of okadaic acid in shellfish, which belongs to the technical field of electrochemical detection. The electrochemical detection method comprises the following steps: firstly, electro-polymerizing a thionine-methylene blue composite film on the surface of a bare gold electrode, and sequentially modifying nanogold and an antibody on the electrode surface through electrode surface modification technology to prepare an electrochemical immunosensor; and then, applying the prepared electrochemical immunosensor to electrochemical detection of okadaic acid in shellfish. A peak current signal is amplified by means of a synergistic effect of the thionine-methylene blue composite polymer film and the nanogold to improve the sensitivity of the sensor; the okadaic acid with special molecular structure is specifically combined with the antibody to form an immune product existing in the form of anion, and the immune product can block the electron transfer property of the electrode surface to achieve non-labeled electrochemical detection. The method disclosed by the invention is used for measuring okadaic acid in shellfish samples, the detection is fast, the sensitivity is high and the result is accurate and reliable.

The invention belongs to the field of immunoassay technical methods, and particularly relates to fluorescence quenching test paper for detecting field okadaic acid as well as a preparation method andapplication of the fluorescence quenching test paper. The fluorescence quenching test paper comprises a sample pad, a combination pad, a nitrocellulose membrane, a water absorption pad and a PVC bottom plate. An OA monoclonal antibody marked by colloidal gold particles is sprayed on the combination pad, detection lines on the nitrocellulose membrane are fluorescent microspheres and OA-BSA mixed solution, and quality control lines are fluorescent microspheres. The fluorescence quenching test paper is high in specificity and free of cross reaction, the sensitivity of the fluorescence quenching test paper is 1.56 ppb and is improved by 9.6 times compared with that of colloidal gold test paper with the same parameters, the detection limit of the fluorescence quenching test paper is 3.12-50 ppb, and the detection time is as short as 9 min. The obtained fluorescence quenching test paper has very high stability on interference of shellfish tissue, can achieve quantification of OA toxin by matching with a simple fluorescence immunochromatographic camera and data analysis software, result judgment is visual, and cause misjudgment of non-professionals can be provided. The fluorescence quenching test paper has bright development and application prospects.

The invention provides an ELISA detection method of red-tide algae toxin okadaic acid (OA) and has the advantages of quickly, simple and sensitive detection of the OA content and low cost. The ELISA detection method of the invention comprises the following steps: (1) diluting coated antigens by using carbonate buffer solution to 1:2000, coating ELISA plates; (2) washing the plates for three times and sealing the plates; (3) dumping the blocking solution, washing the plates, separately stirring and mixing OA standard solution which is diluted to series concentrations and shellfish extract liquid with isometric anti-OA monoclonal antibody which is diluted to 1:10,000 evenly to react; (4) dumping the residual liquid, washing the plates, adding goat anti-rat IgG antibody labeled with HRP, diluting the solution to 1:5000 with each pore containing 100mu l of solution, keeping temperature for 1h at 37 DEG C; (5) dumping the residual liquid, washing the plates, adding developer and measuring the OD450 value of each pore.

The present invention offers the monoclonal antibodies that have a high specificity to ciguatoxins, which are the causative poisons of the ciguatera poisoning. The monoclonal antibodies were produced using synthetic hapten conjugates as artificial antigens, which were prepared by the conjugation of a synthesized hapten consisting of the IJKLM-ring fragment of ciguatoxins with proteins such as KLH, BSA, etc. The monoclonal antibodies show a specific reactivity to ciguatoxins, but little or almost no reactivity to other marine polyether toxins such as the okadaic acid, etc.

The invention discloses an application of the phosphatase inhibitor Okadaic acid, OA in preparing drug used to prevent partial fibrosis and cicatrisation after the glaucoma folliculus operation, preparing medical composite with effect amount Okadaic acid, OA with medical findings, employing water, normal saline or phosphate buffer solution as solvent, with the proper concentration being 50-200 nmol / l, daubing or spraying the product on the folliculus part, or eye dropping before operation (two weeks).

The invention discloses a quick detection kit for okadaic acid toxins in shellfish, belonging to the technical field of food detection. The quick detection kit comprises (1) protein phosphatase 2A freeze-dried powder, (2) an okadaic acid concentration gradient series standard solution, (3) a buffer solution, (4) a protein phosphatase 2A dilute solution, (5) a substrate developing solution, and (6) a high-concentration marking solution. According to the technical scheme disclosed by the invention, the quick detection kit inherits the advantage that a mouse bioassay can establish a dosage-effect relation, so that the relative toxicity of the toxins can be directly reflected; compared with the mouse bioassay, the quick detection kit has the advantage that a large batch of samples (84 samples can be detected within 4 hours) can be detected within short time. The detection limit of the method disclosed by the invention is 50 microgram OA eq. / kg shellfish tissue; compared with the detection limit, 200 microgram OA eq. / kg shellfish tissue, of the conventional mouse bioassay, the detection limit of the method disclosed by the invention is greatly reduced; meanwhile, the ethic problem of an animal experiment is solved. The kit disclosed by the invention is low in cost and easy to operate.

The invention relates to the detection device of detecting the red tide toxin. An immune colloidal gold test paper to detect the lienteric toxin includes the sample pad, colloidal gold pad, NC film, the suction pad and PVC backing. The PVC backing is adhibited with the sample pad, colloidal gold pad, NC film and the suction pad. The colloidal gold pad is covered with the monoclonal antibody against okadaic acid marked by colloidal gold. The NC membrane is encrusted with the detection line composed by the okadaic acid and ovalbumin and quality controlling line composed by the multi antibody of sheep against rat. The test paper has high specialty, detection sensitivity, quick detection and simple pretreatment character. It needs not any device and convenience to carry and has low detection cost; the test paper has good stability and can be stored for six months in room temperature; the detection result has high veracity and precision.

The invention belongs to the technical field of aptamer design, and particularly relates to an aptamer design method based on single-nucleotide molecular docking. Starting from an experimental structure of a target small molecule, positions of nucleotides combined around the target small molecule are obtained through hydration docking; then, the discrete nucleotides are assembled into a complete aptamer; and finally, the binding capacity of the designed aptamer and the target small molecule is detected by adopting an MST experiment. The aptamer design method in the invention has been used in the design of aptamers of toxin small molecule okadaic acid (OA); the toxin is a widely distributed marine biological toxin, and can cause diarrhea type shellfish poisoning, so that the detection of OAin marine products is of great significance to food safety; an aptamer OA-D1 targeting OA is obtained by design through a calculation method; the MST experiment verifies that the equilibrium dissociation constant of the aptamer and OA is 75.6 nM; and therefore, the calculation design method is reasonable and effective.

The invention discloses application of okadaic acid in preparing a drug composition for treating periprosthetical osteolysis. The okadaic acid is of a structure shown as a formula (I). By studying treatment effect of the okadaic acid on periprosthetical osteolysis, a novel path is provided for preventing and treating periprosthetical osteolysis.

The invention relates to a magnetic-nanoparticle-modification-based enzyme sensor used for detecting okadaic acid. The enzyme sensor comprises an insulated base body, and an electrode system including a work electrode, a reference electrode and a counter electrode, which is formed on the base body, wherein a biological enzyme layer is fixed on the work electrode, and comprises magnetic nanoparticle modified protein phosphatase 2A (PP2A) and an electronic transfer body, the diameters of magnetic nanoparticles are 250nm-350nm, and the invention also relates to a preparation method of the enzyme sensor. Compared with a common enzyme sensor, the enzyme sensor has the advantages that the loading rate of PP2A on the surface of the work electrode is improved through the magnetic nanoparticle modified PP2A, so that the current response value of the enzyme sensor is improved by 10 times or more compared with the common PP2A sensor, and when the enzyme sensor is used for detecting okadaic acid of an aquatic product, the detection limit is lower than 0.20microgram / L, the reproducibility RSD is smaller than 5%, and the detection time consumption is smaller than 0.5h.

The invention discloses an Okadaic acid detection kit, and belongs to the technical field of detection kits in laboratories. The Okadaic acid detection kit comprises four hemispheric grooves, a flip cover, a square hole, a square buckle, hand ropes, side plates, a zipper, a spring partition plate, a vibration absorption spring, a vehicle wheel frame, wheels, a bottom plate, a support, a cylinder pin, a vibration damper plate, a downward-concaved bottle position, a honeycomb-shaped buffering paperboard support, a 96-hole elisa plate, a plate and a 12-hole batten. The front, back, left and right of the bottom plate are respectively connected with one side plate, the back side plate is connected with the flip cover, the four hemispheric grooves are formed in the flip cover, the square hole is formed in the flip cover, the square buckle is arranged on the front side plate, and the hand ropes are arranged on the left side plate and the right side plate. A reagent bottle is placed and gotten conveniently, and the safety of reagent bottle transportation is guaranteed through the buffering device around the reagent bottle.

The invention discloses a fluorescent nucleic acid aptamer sensor for detecting okadaic acid, a preparation method of the fluorescent nucleic acid aptamer sensor and a method for detecting okadaic acid by using the fluorescent nucleic acid aptamer sensor, and belongs to the technical field of harmful substance detection. According to the invention, the fluorescent nucleic acid aptamer sensor for detecting the okadaic acid is formed by self-assembly of a nucleic acid aptamer which is marked by a fluorophore at the 5' end and can specifically recognize the okadaic acid and graphene oxide. When the fluorescent nucleic acid aptamer sensor is used for detecting okadaic acid, the operation steps are simple and clear, the cost is low, the specificity is good, the detection range is between 0.5 ng / mL and 200 ng / mL, and the sensitivity is high.

The invention discloses a preparation method and application of amino modified Fe3O4 nano microspheres. By taking ferric trichloride hexahydrate, anhydrous sodium acetate, polyethylene glycol, cysteamine hydrochloride (CSH) and ethylene glycol as raw materials, the amino-modified Fe3O4 nano microspheres (Fe3O4-NH2) are prepared. Okadaic acid in scallops is extracted by using the Fe3O4-NH2 as an adsorbent. Extraction conditions of the Fe3O4-NH2 on okadaic acid are optimized; and when the pH value is 7, the mass of the adsorbent is 20 mg, the vortex adsorption time is 20 min, the eluent is a mixture of ammonia water and methanol in a ratio of 10%: 90%, and the volume of the eluent is 4 mL, the extraction efficiency of the Fe3O4-NH2 on the okadaic acid in shellfish is optimal.

The invention relates to a biosensor for detecting okadaic acid. The biosensor comprises an insulated substrate and an electrode system, wherein the electrode system is formed on the substrate and comprises a working electrode, a reference electrode and a counter electrode; and a biological enzyme layer is fixed on the working electrode and comprises protein phosphatase 2A (PP2A enzyme) and an electron transport complex. The invention further relates to a preparation method of the biosensor, and mainly relates to fixation of the PP2A enzyme. Compared with the prior art, the biosensor taking the PP2A enzyme as a fixed enzyme layer is prepared by utilizing the inhibition effect of okadaic acid to the activity of the PP2A enzyme; the content of okadaic acid in a sample is determined according to the activity inhibiting degree of the PP2A enzyme, namely, the concentration of okadaic acid in the sample is converted into an electric signal to be detected, and therefore, the detection result is visual; simultaneously, by virtue of the specificity and the high efficiency of PP2A enzyme catalytic reaction, the detection process is high in specificity, the accuracy rate of a detection structure is high, the detection period is short, and instant and rapid detection requirements can be met.

The invention relates to a chemiluminesent immunoassay kit for detecting okadaic acid, belonging to the technical field of laboratory detection kits. The chemiluminesent immunoassay kit comprises a latching head, a kit cover, a kit body, a cut sponge, a buckling groove, a drawer, an overturning cover, an elisa (enzyme-linked immuno sorbent assay) plate support, cylindrical pins, a support, a reagent bottle support, a groove and a 96-well elisa plate, wherein the latching head is arranged on the kit cover which is connected with the kit body, the buckling groove is arranged on the kit body, the drawer is arranged under the kit body, the elisa plate support and the reagent bottle support are arranged in the kit body, the overturning cover is connected with the elisa plate support, the cut sponge is arranged on the bottle surface inside the elisa plate support, the 96-well elisa plate is arranged on the cut sponge in the elisa plate support, cylindrical pins are arranged on left and right side surfaces of the elisa plate support, the reagent bottle support is arranged under the elisa plate support, cylindrical pins are arranged on left and right side surfaces of the reagent bottle support, the folding design of the elisa plate support and the reagent bottle support can achieve firmness between the elisa plate support and the reagent bottle support and ensures that a reagent bottle can be taken conveniently.

The invention discloses a gold magnetic nanoprobe based on ordered arrangement of aptamers and application of the gold magnetic nanoprobe in okadaic acid detection (OA). The gold magnetic nanoprobe is formed by combining nanogold spherical nucleic acid and nano magnetic beads, wherein nucleic acid sequences are orderly arranged on the surface of the nanogold spherical nucleic acid; the nucleic acid sequence is formed by an aptamer with an adenine base polymeric chain at the 5'end and a DNA complementary chain with a fluorescent label at the 5 'end. The surface of the gold magnetic nanoprobe is modified with high density and is loaded with orderly arranged aptamers, so that the gold magnetic nanoprobe is high in specific recognition efficiency and simple and convenient to separate a target object, and can react with okadaic acid to release a fluorescently-labeled DNA complementary chain, and laser-induced fluorescence detection is applied, so that high-sensitivity analysis of trace OA is realized.

The invention discloses a preparation method of a bamboo-like nitrogen / metal cobalt doped carbon nanotube material as a magnetic solid-phase extraction adsorbent and application of the bamboo-like nitrogen / metal cobalt doped carbon nanotube material in adsorption and enrichment of okadaic acid. The specific surface area of the bamboo-like nitrogen / metal cobalt doped carbon nanotube material disclosed by the invention reaches 154 cm<3> / g; the nitrogen element is doped, so that the material has a large number of adsorption sites; [pi] electrons on the carbon nanotubes and [pi] electrons on the okadaic acid can form pi-pi conjugation; the nitrogen element and the okadaic acid can adsorb the okadaic acid through hydrogen-bond interaction. In addition, the bamboo-like structure enables the okadaic acid to have excellent structural stability, thermal stability and chemical stability, and meanwhile, the okadaic acid has the characteristic of magnetism and is also beneficial to adsorption of subsequent separation work. When the adsorbent is applied to adsorption and enrichment of okadaic acid in the okadaic field, the effects of rapid adsorption (5 min) and rapid desorption (15 min) can beachieved.

The invention provides a paper-based coupling enhanced Raman (SERS) sensor based on aptamer specific capture performance and a preparation method thereof, and provides high-specificity analysis of okadaic acid in marine water and marine products by means of the paper-based coupling enhanced Raman sensor. The application of the paper-based coupling enhanced Raman sensor is implemented by the stepsof: assembling dens and ordered nano gold particles on the surface of paper fibers in a reduction induced mode so as to construct a surface plasma resonance nano gold paper SERS substrate; synthesizing a three-dimensional space dendritic 3D silver material in situ, so as to construct a high-hotspot-density electromagnetic field enhanced SERS coupling double-layer; and introducing complementary DNAchains to establish a specific capture-surface coupling enhanced Raman analysis interface, so as to realize rapid and precise Raman spectrum analysis. The purpose of field detection is achieved, thepreparation method is simple, the cost is low, large-scale preparation can be achieved, and theoretical and technical support is provided for seawater treatment.

The invention discloses a preparation method of an okadaic acid toxin molecularly imprinted-quantum dot polymer and an application. The preparation method comprises the steps of taking okadaic acid as a template molecule and adding a quantum dot fluorescent nanomaterial; and initializing polymerization in the presence of a cross-linking agent tetraethylortho silicate and a functional monomer, and then removing the template molecule in the obtained polymer by adopting an ultrasonic-assisted extraction method to obtain the okadaic acid toxin molecularly imprinted-quantum dot polymer. The application method comprises the steps of sampling a sample extract solution to an okadaic acid Spin solid-phase extraction column; centrifuging, collecting an eluent and filtering the eluent through a 0.22-micron filter membrane; and measuring the content of okadaic acid toxin by adopting an LC-MS / MS. The method has the advantages of being high in sensitivity, good in stability, high in selectivity, simple in method and convenient to operate, and the targets of simply and efficiently achieving selective separation, purification an enrichment of okadaic acid saxitoxin in a shell sample are achieved.

The invention relates to a preparation method of an okadaic acid three-dimensional gold nanopillar array immunity electrode. The preparation method comprises the following steps: depositing gold on apolycarbonate filter membrane with the aperture being 80-200 nm with a chemical deposition-electro-deposition method, so as to obtain a three-dimensional gold nanopillar array electrode; electro-polymerizing and decorating poly(thionine) on the surface of the three-dimensional gold nanopillar array electrode with a cyclic voltammetry, so as to form a poly(thionine) / three-dimensional gold nanopillar array electrode; binding glutaraldehyde on the poly(thionine) / three-dimensional gold nanopillar array electrode and fixing an okadaic acidantibody on the glutaraldehyde, so as to form an okadaic acidantibody / glutaraldehyde / poly(thionine) / three-dimensional gold nanopillar array electrode; and finally, closing the electrode with bovine serum albumin, so as to obtain the okadaic acid three-dimensional gold nanopillar array immunity electrode. The electrode provided by the invention has a three-dimensional structure and is large in superficial area, simple in preparation, high in stability, firmin fixation of the antibody, simple and convenient in operation, low in detection limit and high in sensibility, and rapid detection can be achieved.

The invention particularly relates to a method for establishing a zebrafish senile dementia model and application. A glucose culture solution and an okada acid culture solution are included,the glucose culture solution and the okada acid culture solution are alternately used for culturing zebrafishes, and a single culture time of the glucose culture solution orthe okada acid culture solution is 22-26 hours. The method disclosedis the first methodwhich alternately cultures the zebrafishes with a specific concentration of the glucose culture solution and the okada acid culture solution, zebrafish senile dementia-like symptoms can be quickly and effectively caused, thereby greatly reducing the dosage of okada acid, and experimental resources are saved.

The invention relates to the technical field of medicine preparation, particularly to an okadaic acid derivative and a preparation method thereof. According to the invention, the okadaic acid derivative prepared by the preparation method is high in yield and novel and coordination compound in structure; compared with the method in the prior art, the preparation method adopted by the invention is simple and practical, has few procedures, reduces the generation of byproducts, and improves the purity of the prepared okadaic acid derivative; and the okadaic acid derivative has potential application and wide market prospect in the field of pharmacy.

The invention relates to a chemiluminesent immunoassay kit for detecting okadaic acid, belonging to the technical field of laboratory detection kits. The chemiluminesent immunoassay kit comprises a latching head, a kit cover, a kit body, a cut sponge, a buckling groove, a drawer, an overturning cover, an elisa (enzyme-linked immuno sorbent assay) plate support, cylindrical pins, a support, a reagent bottle support, a groove and a 96-well elisa plate, wherein the latching head is arranged on the kit cover which is connected with the kit body, the buckling groove is arranged on the kit body, the drawer is arranged under the kit body, the elisa plate support and the reagent bottle support are arranged in the kit body, the overturning cover is connected with the elisa plate support, the cut sponge is arranged on the bottle surface inside the elisa plate support, the 96-well elisa plate is arranged on the cut sponge in the elisa plate support, cylindrical pins are arranged on left and right side surfaces of the elisa plate support, the reagent bottle support is arranged under the elisa plate support, cylindrical pins are arranged on left and right side surfaces of the reagent bottle support, the folding design of the elisa plate support and the reagent bottle support can achieve firmness between the elisa plate support and the reagent bottle support and ensures that a reagent bottle can be taken conveniently.

The invention particularly relates to a soaking solution for reducing the content of an okadaic acid toxin. To overcome the shortcomings in a technical measure of reducing the okadaic acid toxin in meat at the present, the technical scheme of the invention is that the soaking solution for reducing the content of the okadaic acid toxin is characterized by being prepared from the following components in parts by weight: 10 to 100 parts of gingerol, 10 to 100 parts of taraxacin, 10 to 100 parts of tea polyphenol, 100 to 1,000 parts of ethanoland 500 to 1,000 parts of water. The component constituents of the soaking solution are simple and easy to obtain; a soaking method is simple; after shell meat is treated, the nutritional components of the shell meat cannot be lost, and the meat cannot beinjured; and meanwhile, the content of the okadaic acid toxin can be effectively reduced.