47 results about "Sterigmatocystin" patented technology

The invention provides a preparation method for a fluorescence sensing material based on molecular imprinting and carbon dots. The method comprises the following steps: adding a substitute template and a functional monomer into a mixed solution composed of chloroform, acetonitrile and toluene, carrying out stirring and allowing the mixed solution to fully dissolve and react, then adding the carbon dots, carrying out stirring and then adding a crosslinking agent and an initiator, carrying out ultrasound and then introducing nitrogen for water bath and incubation, carrying out filtering, carrying out placing in a vacuum drying oven for aging, carrying out washing with an extracting solvent so as to remove template molecules, and carrying out vacuum drying so as to obtain a molecular imprinting and carbon dot fluorescent sensing material. The preparation method for the fluorescence sensing material provided by the invention has the advantages of high stability, light-free bleaching, low toxicity and capability of directly forming a sensing material by embedding into molecular imprinting polymer through polymerization reaction without the need of subsequent modification; and the sensing material in the invention is simple in preparation process, and has specific recognition effect of the molecular imprinting polymer, strong fluorescence characteristic of the carbon dots, and selective recognition effect to sterigmatocystin.

The invention discloses an immunosorbent comprising a solid-phase carrier and an antibody coupled with the solid-phase carrier. The antibody comprises a monoclonal antibody secreted by a hybridoma cell line CGMCC NO.5504 and a monoclonal antibody secreted by a hybridoma cell line CGMCC NO.5505. The invention also provides a kit comprising the immunosorbent or an immunoaffinity column. Also, the invention provides applications of the immunosorbent, immunoaffinity column, and kit in detecting aflatoxins, sterigmatocystin, zearalenone congeners and ochratoxin A. The invention specifically provides separation and detection methods. According to the invention, the specific monoclonal antibodies with stable performances are developed, such that simultaneous purification and detection of 6 aflatoxins, sterigmatocystin, 6 zearalenone congeners and ochratoxin A are realized.

The invention relates to the field of biotechnology and provides a multiple PCR detection primer group, a kit and a detection method for fungus producing aflatoxin. According to the invention, through combined utilization of a fungus universal molecular marker sequence (internal transcribed spacer, ITS) and three key genes, comprising an aflatoxin production regulating gene (aflR), a sterigmatocystin-o-methoxy transferase gene (omt-1), and versicolorin A dehydrogenase gene (ver-1), in the biochemically synthetic process of the aflatoxin, the multiple PCR detection primer group, the kit and the detection method have the technical effects of wide coverage range, strong specificity and high sensitivity during the detection of the fungus producing aflatoxin. The primer group, the kit and the detection method are short in detection time, are simple in operation and are low in device requirement, can save large amount of manpower, materials and costs, are suitable for quick detection and are easy to promote in practical production.

The invention relates to an immunoadsorbent for purifying five kinds of mycotoxins including fumonisin B₁ and aflatoxin B₁, and a complex affinity column. The immunoadsorbent comprises a solid-phase support, and an anti-fumonisin B₁ monoclonal antibody, an anti-aflatoxin B₁ monoclonal antibody, an anti-ochratoxin A monoclonal antibody, an anti-zearalenone monoclonal antibody and an anti-sterigmatocystin monoclonal antibody which are coupled to the solid-phase support, wherein the anti-fumonisin B₁ monoclonal antibody is secreted by a hybridoma cell strain Fm7A11, and the hybridoma cell strain Fm7A11 has been preserved at China Center for Type Culture Collection, Wuhan University, Wuhan, China on Mar. 29, 2016 with the preservation number of CCTCC No. C201636. The complex affinity column can be used for purification and detection of a fumonisin B₁ sample, an aflatoxin B₁ sample, an ochratoxin A sample, a zearalenone sample and a sterigmatocystin sample at the same time.

The invention discloses a biosensor for rapidly detecting sterigmatocystin and an assembling method thereof; the biosensor comprises a substrate electrode and a reaction layer assembled on the substrate electrode, the reaction layer comprises a polymer film, an electronic carrier, chitosan gel and 6-methoxy group furocoumarin coumarin oxidase, the 6-methoxy group furocoumarin coumarin oxidase is fixed in the electronic carrier-chitosan mixing membrane, and then is assembled on the substrate electrode decorated by the polymer film, thereby obtaining the biosensor for detecting the sterigmatocystin; the biosensor has high sensor sensitivity, rapid response time and strong anti-jamming capability, the detection lower limit to the sterigmatocystin can reach 3ng/ml, the biosensor can be used for practical sample detection, and the average recovery rate is 95.8 percent; the biosensor has simple manufacturing, convenient usage, good stability and long service life and provides a new detection method for rapidly detecting the sterigmatocystin content in practical samples.

The invention discloses an anti-sterigmatocystin monoclonal antibody. The antibody comprises a light chain and a heavy chain, wherein an amino acid sequence of a variable region of the light chain is shown in a sequence table SEQ ID NO. 1, and an amino acid sequence of a variable region of the heavy chain is shown in a sequence table SEQ ID NO. 2. The invention further discloses a method for preparing the antibody. A detection kit for detecting sterigmatocystin is prepared by using the antibody disclosed by the invention. The sensitivity of the kit reaches 10 pgs per microliter, so that the content of the sterigmatocystin in a sample can be effectively detected. The kit is convenient to use and does not need valuable instruments, thereby facilitating large-scale popularization and application.

The invention relates to the field of biotechnology and provides a single PCR detection primer pair group aiming to a fungus producing aflatoxin, a detection method, and an application thereof in detection on the fungus producing aflatoxin. In the invention, the single PCR detection primer pair group is composed of primer pairs that aim to a fungus universal molecular marker sequence (internal transcribed spacer, ITS) and three key genes, comprising an aflatoxin production regulating gene (aflR), a sterigmatocystin-o-methoxy transferase gene (omt-1), and versicolorin A dehydrogenase gene (ver-1), in the biochemically synthetic process of the aflatoxin. Through the primer pair group, technical effects of wide coverage range, strong specificity and high sensitivity during the detection of the fungus producing aflatoxin are achieved. The primer pair group is short in detection time, is simple in operation and is low in device requirement, can save large amount of manpower, materials and costs, is suitable for quick detection and is easy to promote in practical production.

The invention discloses a hybridoma cell strain CGMCC NO. 5506 and a monoclonal antibody obtained by secretion of the cell strain. The invention also provides an immunoadsorbent comprising a solid phase vector and an antibody coupled with the solid phase vector, wherein the antibody is the above monoclonal antibody; and an immunoaffinity column loaded with the immunoadsorbent. The invention also provides a kit containing the above immunoadsorbent or the above immunoaffinity column, and applications of the above immunoadsorbent, the above immunoaffinity column and the kit in detecting aflatoxins and sterigmatocystins. Besides, the invention provides a separation method and a detection method specifically. The monoclonal antibody with stable and specific performance is developed by the invention, so that purification and detection for six aflatoxins and sterigmatocystins at the same time are realized.

The invention discloses an immunochromatographic time resolved fluorescence kit for synchronously detecting aflatoxins, and an application of the kit. The kit comprises an immunochromatographic time resolved fluorescence test strip and a sample reaction bottle containing europium-labeled monoclonal antibody freeze-dried samples, wherein the immunochromatographic time resolved fluorescence test strip comprises a bottom plate; a water absorption pad, a detection pad and a sample pad are adhered to the adhesive surface of the bottom plate in sequence from top to bottom; the adjacent pads are connected in an overlapped mode at the joint; the detection pad takes a nitrocellulose membrane as a base pad; a transverse quality control line and a detection line are arranged on the nitrocellulose membrane from top to bottom; the quality control line is coated with a rabbit anti-mouse polyclonal antibody; the detection line is coated with protein conjugates of the toxins; and an anti-cyclopiazonicacid monoclonal antibody is secreted by a hybridoma cell strain YTT-2 with a collection number of CCTCC NO.C201871. The kit can be used for synchronously detecting the content of the aflatoxins, sterigmatocystins and cyclopiazonic acids in the samples, and has the characteristics of simplicity in operation, quickness and high sensitivity.

The invention discloses a time-resolved fluorescence kit for detecting diacetoxyscirpenol, aflatoxin B1 and sterigmatocystin simultaneously. The time-resolved fluorescence kit comprises an immunochromatographic time-resolved fluorescent test strip and a sample reaction bottle containing an europium-marked monoclonal antibody resisting diacetoxyscirpenol, aflatoxin B1 and sterigmatocystin, whereina water absorption pad, a test pad and a sample pad are arranged at one side of the immunochromatographic time-resolved fluorescent test strip in sequence from top to bottom, adjacent pads overlap andare connected at the joint, the detection pad uses a nitrocellulose membrane as a base pad, a quality control line and three detection lines are transversely arranged on the nitrocellulose membrane from top to bottom, the quality control line wraps a rabbit anti-mouse polyclonal antibody, and the three detection lines respectively wrap toxin-protein conjugates respectively. The kit can rapidly detect the three fungal toxins like diacetoxyscirpenol, aflatoxin B1 and sterigmatocystin simultaneously.

The invention discloses a colloidal gold immunoassay test strip for synchronously detecting aspergillus flavus fungaltoxin as well as preparation and application thereof. The colloidal gold immunoassay test strip comprises a bottom plate, wherein an absorbent pad, a detection pad, a gold label pad and a sample pad are stuck to one side of the bottom plate from top to bottom in sequence; adjacent pads are connected at joints in an overlapping manner; the detection pad takes a nitrocellulose membrane as a base pad; a quality control line and detection lines are transversely arranged on the nitrocellulose membrane from top to bottom; three detection lines are positioned below the quality control line, and distributed at intervals; the three detection lines are coated with toxin protein conjugates respectively; the quality control line is coated with a rabbit anti-mouse polyclonal antibody; and an anti-cyclopianic acid monoclonal antibody is generated by secretion of a hybridoma cell strain YTT-2 of which the collection number is CCTCC NO.C201871. The colloidal gold immunoassay test strip can be applied to the synchronous detection of aflatoxin, sterigmatocystin and cyclopiazonic acid,is simple and rapid in operation, and has high sensitivity.

The invention relates to a non-toxic aspergillus flavus strain and application thereof. A non-toxic aspergillus flavus XZCY1805 disclosed by the invention does not produce various toxins such as aflatoxin (AFT), cyclopiamic acid (CPA), aspergillus flavus tremorine (Aflatrem), sterigmatocystin (ST) and the like, and is a biocontrol strain with high safety and strong competitive advantage. The non-toxic aspergillus flavus XZCY1805 disclosed by the invention has a strong toxic inhibition effect on a standard toxic aspergillus flavus strain 3.4408 and toxic aspergillus flavus separated from different peanut production places, and can remarkably reduce the contents of the aflatoxin, the cyclopiamic acid, the aspergillus flavus tremorin and the sterigmatocystin in agricultural products such as peanuts and the like.

The invention discloses a multiplex PCR primer combination, a multiplex PCR kit and a detection method for rapidly detecting aflatoxin-producing bacteria. The multiplex PCR primer combination comprises a primer pair, wherein the primer pair is used for specifically amplifying sterigmatocystin A dehydrogenase gene ver-1, and comprises an upstream primer with a nucleotide sequence as shown in SEQ ID No.1 and a downstream primer with a nucleotide sequence as shown in SEQ ID No.2; a primer pair used for specifically amplifying the sterigmatocystin B gene verB and comprises an upstream primer with a nucleotide sequence as shown in SEQ ID No.3 and a downstream primer with a nucleotide sequence as shown in SEQ ID No.4; and a primer pair used for specifically amplifying the ITS sequence of the fungus. The multiplex PCR primer combination disclosed by the invention is strong in specificity and high in sensitivity, and does not interfere with each other.

The invention discloses multi-PCR primers, method and kit for simultaneously detecting four kinds of enterotoxigenic fungi of pollution of traditional Chinese medicinal materials. The four kinds of enterotoxigenic fungi are an aflatoxin production bacterium, a sterigmatocystin production bacterium and an ochratoxin production bacterium. Based on the multi-PCR detection method, the multi-PCR primers comprise a first primer pair, a second primer pair, a third primer pair and a fourth primer pair which are subjected to the PCR reaction in the same reaction tube. The multi-PCR system capable of directly detecting the enterotoxigenic fungi of pollution of traditional Chinese medicinal materials is constructed without separation and purification, all the primers are high in specificity, the detection limit of the method is 102-103 CFU/g, and the detection sensitivity is high.

The invention provides non-toxin-producing aspergillus flavus SX0104 and application thereof, and particularly relates to the technical field of microorganisms. According to the provided non-toxin-producing aspergillus flavus SX0104, the preservation number of the aspergillus flavus SX0104 is CCTCC NO:M2020520. The aspergillus flavus SX0104 does not produce aflatoxin, aspergillus flavus tremorin or sterigmatocystin, is a biocontrol strain with high safety and strong reproductive capacity, and has the capabilities of efficiently and competitively inhibiting the growth, reproduction and toxin production of toxin-producing aspergillus flavus; and the content of the aflatoxin, the aspergillus flavus tremorin and the sterigmatocystin in agricultural products such as peanuts can be obviously reduced.

The invention discloses an ELISA (enzyme-linked immuno sorbent assay) kit for detecting sterigmatocystin, wherein the kit comprises a sterigmatocystin antibody, the antibody comprises a light chain and a heavy chain, the amino acid sequence of the light chain is shown as the sequence table SEQ ID NO.1; and the amino acid sequence of the heavy chain is shown as the sequence table SEQ ID NO.2. The invention further discloses a method for preparing the antibody. The ELISA kit provided by the invention comprises a horse radish Peroxidase substrate buffer liquid, a protein standard product ST (100 micro g/ml, 0.1ml), dox-ova BSA-ST and negative control sample BSA. The invention also discloses a using method of the kit.

The invention discloses a preparation method of a reagent plate for detecting sterigmatocystin. A colloidal gold binding pad of a nitrocellulose film is coated with an anti-sterigmatocystin monoclonal antibody-colloidal gold mark, a detection line and a quality control line are orderly arranged from a sample pad to a water absorption pad and are respectively coated with a sterigmatocystin-carrier protein conjugate and goat anti-mouse IgG, and the carrier protein is a rabbit serum protein. The nitrocellulose film is spray-coated with the sterigmatocystin-carrier protein conjugate and goat anti-mouse IgG through a film formation machine so that the detection line and the quality control line are obtained, the nitrocellulose film is dried by an oven at a temperature of 38 DEG C for 8h, the colloidal gold binding pad is spray-coated with the anti-sterigmatocystin monoclonal antibody-colloidal gold mark through the film formation machine, and the colloidal gold binding pad is dried by the oven at a temperature of 38 DEG C for 8h. The reagent plate has the advantages of fast detection rate, sensitivity, accuracy, quantification and simple operation. The preparation method has simple processes, realizes detection of a single sample, has a low production cost and greatly reduces a detection cost.

The invention belongs to the field of food detection methods, and discloses a method for detecting fungaltoxin in spices and products thereof. A liquid chromatogram-tandem mass spectrum/mass spectrum determination method is used for determining the content of aflatoxin B1, aflatoxin B2, aflatoxin G1, aflatoxin G2, ochratoxin A, fumonisins B1, fumonisins B2, deoxynivalenol, fumonisins B1, fumonisins B2, deoxynivalenol, fumonisins B1, fumonisins B2, fumonisins B1, fumonisins B2, fumonisins B1, fumonisins B2, fumonisins B1, fumonisins B1, fumonisins B2, fumonisins B1, fumonisins B2, fumonisins B1, fumonisins The medicine is prepared from the following raw materials: 3-acetyl deoxynivalenol, 3-acetyl deoxynivalenol, 15-acetyl deoxynivalenol, T-2 toxin, HT-2 toxin, zearalenone and sterigmatocystin. By setting the mass spectrum condition and liquid phase condition of each mycotoxin, various mycotoxins in spices and products thereof can be effectively extracted, purified, separated and detected, the experiment precision and detection limit both meet the experiment requirements, and simultaneous detection of 14 mycotoxins is realized.

The invention relates to aspergillus versicolor ZJUTE2 and application of the aspergillus versicolor ZJUTE2 in fermentation preparation of sterigmatocystin. The invention reports a method for preparing high-purity ST with high yield through fermentation culture of the strain for the first time, so that the yield of ST reaches 2.04 g/kg rice, and the purity reaches 99.41%; the preparation method has the advantages of simple fermentation conditions, easy culture of strains, few process steps, short production period and the like, and has the potential of industrial and large-scale production. The method has important significance for solving the industrial problem that the high-purity ST standard substance or reference substance depends on import for a long time.