133 results about "Fumonisin" patented technology

The invention relates to an electrochemical lateral-flow immunity quantitative test paper sensor based on microclearance array electrode, and a method thereof used for detecting biological toxins such as ochratoxin A and fumonisin B1. The sensor includes two parts, namely, immune chromatographic analysis test paper strip and electrochemical detecting part. The fast detecting test paper sensor has strong specificity, can realize quantitative detection, can be used at the temperature between 4 and 40 DEG C, and the result can be observed after ten minutes, thus being suitable for units or individuals to quickly detect ochratoxin A and fumonisin B1 in animal derived food samples, and being expected to become an effective technical means for the field screening of ochratoxin A and fumonisin B1 in food and feed samples.

The invention relates to a hybridoma cell line and application thereof. The hybridoma cell line can generate anti-fumonisin FB1 monoclonal antibodies, and is used for detecting the pollution condition of fumonisin FB1. The hybridoma cell line is characterized in that: the collection number of the hybridoma cell line is CCTCC NO: C201008. The invention has the advantages that: the cell line can specifically secrete the monoclonal antibodies which aim at the fumonisin FB1, and has high sensitivity and specificity; and the antibodies generated by the cell line can be applied to various ways, andhave high application value in practical production.

The invention relates to a method for detecting fumonisins based on immuno-magnetic bead combined enzyme-linked immunosorbent assay, belonging to the technical field of chemical detection. The method comprises the steps of: preparing fumonisins-KLH conjugate and fumonisins-OVA conjugate, combining the fumonisins-KLH conjugate with an immuno-magnetic bead, and preparing a magnetic bead for detecting the fumonisins; then, using the fumonisins-KLH conjugate to prepare fumonisins monoclonal antibody, applying a competition ELISA method for detecting the fumonisins together with the magnetic bead for detecting the fumonisins, and obtaining the magnetic bead for detecting the fumonisins by a magnetic separating method; and developing and obtaining the detection result by an enzyme linked immunosorbent assay. The method is used for the fumonisins sample which is lower than detection limit, and enlarges the combination superficial area by enrichment of the immuno-magnetic bead and the full diffusion of the magnetic bead in the liquid, thus indirectly changing the detection limit, improving the detection sensitivity and avoiding undetected error.

The invention belongs to the technical field of genetic engineering, and in particular relates to screening of a single-chain antibody against fumonisin and application thereof in immunoassay of fumonisin. The screening method comprises the following steps of: directly setting off from coupled antigen FB1-KLH(keyhole limpet hemocyanin) immunized mouse spleen cells, establishing a single-chain antibody gene library by a molecular cloning method and technology, then screening by a phage display technology, expressing ELISA(enzyme-linked immuno sorbent assay) test, sequencing, and finally obtaining the single-chain antibody against fumonisin named as FB-Mu 1H3 with high affinity and a coding gene thereof. The single-chain antibody can be directly applied to assay of fumonisin after being expressed and purified in a large quantity in escherichia coli, and the applications include assay of fumonisin pollution in field crops, feed, grain or food.

The invention discloses a method for detecting fumonisin by a colloidal gold immunochromatographic test in the technical field of biological detection engineering. The method comprises the following steps: conjugating fumonisin hapten with KLH and OVA respectively; preparing a monoclonal antibody of the fumonisin from the obtained fumonisin-KLH conjugate by a conventional method; preparing 40nm colloidal gold by a trisodium citrate reducing method, and labeling the monoclonal antibody of the fumonisin subject to dialysis desalination treatment with the colloidal gold; spraying the obtained monoclonal antibody protein onto a gold-labeled pad, spraying the fumonisin-OVA conjugate onto a T line, spraying a rabbit antimouse monoclonal antibody onto a C line to form a reagent paper tape, and drying; and extracting a sample to be tested with methanol, centrifuging and taking supernatant, adding the supernatant to a sample groove of the reagent paper tape dropwise to acquire development of the T line and the C line, and identifying to obtain results. The method has advantages of high detection specificity, high accuracy, high detection speed and detection time as short as only 5-10min.

A method for simultaneously detecting zearalenone and fumonisins in the technical field of biological detection engineering, comprising the following steps: 1. Prepare conjugates ZEN-BSA and ZEN-OVA; FB1-KLH and FB1-OVA; 2. Prepare anti-ZEN monoclonal antibody and anti-FB1 monoclonal antibody respectively; 3. Prepare colloidal gold, respectively label anti-ZEN monoclonal antibody and anti-FB1 monoclonal antibody; spray the labeled monoclonal antibody onto the gold label pad ; 4. Spotting on the nitrocellulose membrane; 5. Assemble into a test strip; 6. Drop the methanol extraction supernatant of the sample to be tested into the sample groove of the test strip to obtain the points on the nitrocellulose membrane. The color development result of the sample band, the identification, and the result are obtained. The detection object of the method of the present invention is highly targeted and has high accuracy; the detection speed is fast and the required time is short, the loss of manpower and material resources caused by using two detection methods to detect two toxins is reduced, and it is convenient for promotion and application at the grassroots level.

The invention relates to a preparation and detection method for an ELISA kit detecting Fumonisins. The ELISA kit has the characteristics of sensitive, accurate and fast detection, simple operation and strong specificity, and is suitable for detection of a large number of samples. The kit includes: a Fumonisins antigen coated enzyme label plate, a Fumonisins standard substance, a Fumonisins antibody working solution, a Fumonisins enzyme labeled secondary antibody working solution, a substrate solution A, a substrate solution B, a terminating solution, a concentrated sample dilution solution and a concentrated washing solution. The principle of the Fumonisins detection kit is solid-phase indirect competitive enzyme-linked immunosorbent assay reaction. The extracted sample, the enzyme labeled secondary antibody working solution, and the antibody working solution are added into corresponding enzyme labeled holes, after incubation for a period of time, the substrate solution A and the substrate solution B are added into a washing plate, and under the action of enzyme, the holes can present a blue color. Then the terminating solution is added, and the color changes to yellow from blue. The color depth and the content of Fumonisins in the standard substance or the sample are in an inverse proportion relationship. This method can be directly used for detecting Fumonisins in maize.

Relating to the field of removal of fungal toxins, the invention discloses a compound enzyme, an additive, application thereof and a method for removal of fungal toxins. Specifically, the invention relates to a compound enzyme, which contains amidase and esterase and can remove fungal toxins, especially fumonisins, ochratoxins and T2 toxin, additives containing the compound enzyme and applicationthereof in removal of fungal toxins, especially fumonisins, ochratoxins and T2 toxin, and a method for removal of fungal toxins. According to the technical scheme, combined use of the amidase and esterase can achieve simultaneous removal of ochratoxin A, fumonisins and T2 toxin, the removal efficiency is greatly improved than single use of one enzyme, and vomitoxins, aflatoxins and zearalenone toxin can be removed to certain degree.

The invention provides a group of single-stranded DNA nucleic acid aptamers of fumonisin B1. Each aptamer is obtained by carrying out screening, amplification, sequencing, and analysis on affinity and specificity on a random original single-stranded DNA library with the combination of an SELEX (systematic evolution of ligands by exponential enrichment) technology and a magnetic bead with fumonisin B fixed on the surface. As a special efficient distinguishing aptamer of fumonisin B, the group of DNA aptamers provides a new choice for developing a method for replacing the existing methods which are used for detecting fumonisin B by depending on an antibody, and can be used for analyzing and detecting or separating the fumonisin B in the environment of enriched food. The nucleic acid aptamer is small in molecular weight, low in chemical synthesis cost, easy to mark, strong in affinity and specificity, reversible in degeneration renaturation, high in speed, and suitable for repeated use, room-temperature transportation and long-term storage.

Relating to the field of microorganisms, the invention discloses an inoculant, a feed or additive and a removal method for mycotoxins. The invention specifically discloses an inoculant, which includesat least two of mould, yeast and bacteria, wherein the bacteria can be at least one of brevibacterium, acinetobacter, rhodococcus and pseudomonas. Specifically, the inoculant can remove mycotoxins. The invention also discloses application of the inoculant in removal of mycotoxins, an additive containing the inoculant and a removal method for mycotoxins. According to the technical scheme, combineduse of at least two of the yeast, mould and bacteria can achieve simultaneous removal of ochratoxin A, fumonisins and T-2 toxin, and the removal efficiency of ochratoxin A and fumonisins is the highest.

The invention relates to a hybridoma cell strain AFM1B7, a monoclonal antibody thereof and an aflatoxin M1 flow lag immunization time-resolved fluorescence quick test kit. The hybridoma cell strain AFM1B7 is collected in China Center for Type Culture Collection (CCTCC), and the collection number is CCTCC No.C201020. The monoclonal antibody secreted by the hybridoma cell strain AFM1B7 has high sensitivity and good specificity, the 50% inhibition concentration against aflatoxin M1 is 52pg/mL, and the cross reaction rate with aflatoxins B1, B2, G1 and G2, vomitoxin, zearalenone and fumonisin is less than 0.3%. The aflatoxin M1 flow lag immunization time-resolved fluorescence quick test kit prepared from the monoclonal antibody can be used for quantitatively measuring the content of aflatoxin M1, is simple and quick to operate, and has high accuracy.

The invention discloses a method for degrading fumonisins, which uses electron beams to radiate a sample containing the fumonisins to obtain the sample with reduced fumonisins content. The method can directly radiate finished products such as agricultural products, food, and feed, has simple operation and obvious effects, and makes the degradation rate of fumonisins B1 reach 59.03 percent.

Mycotoxin-deactivating aptamers, especially DNA aptamers, bind to mycotoxins in feed and feed ingredients resulting in the reduction or elimination of toxic and carcinogenic effects of mycotoxins. The invention also discloses a composition comprising a mycotoxin-deactivating aptamer, a binding agent, a biotransforming agent and an antioxidant for detoxifying mycotoxins in feeds. In addition, the invention teaches the methods of preparing said mycotoxin-deactivating aptamer-based composition and also the methods of using it as a feed additive. Furthermore, the invention relates to the use of said mycotoxin-deactivator/s alone, or in a composition comprising said aptamers and other mycotoxin-detoxifying agents, in feeds and feed ingredients for detoxifying the major mycotoxins such as aflatoxins, deoxynivalenol, zearalenone, fumonisins and ochratoxin A.

The invention provides a mycotoxin penicillic acid monoclonal antibody and a preparation method thereof. The preparation method comprises the following steps of coupling penicillic acid to carrier protein respectively so as to obtain complete antigen, immunizing a mouse with the complete antigen, adopting a hybridoma technique and obtaining the monoclonal antibody by cell fusion, screening, cloning, culture expansion and an animal in vivo induction ascites method. The obtained antibody has the titer of 1:2.05x10<5>, is of an IgG1 type, has no cross reaction rate with aflatoxin B1, zearalenone, T-2 toxin and Fumonisins, has the affinity constant of 1.54x10<8> L/mol, and can be used for developing ELISA kits for detecting the penicillic acid.

The invention discloses a method for simultaneously detecting zearalenone (ZEN) and fumonisin B1 (FB1), and also discloses an aptamer sensor for simultaneously detecting the zearalenone and the fumonisin. The aptamer sensor is prepared by the following method: (1) covering a glassy carbon electrode with reducing molybdenum disulfide and a gold nanoparticle composite (rMoS2-Au) nanomaterial, and drying; (2) subsequently dripping a mixed solution of a zearalenone nucleic acid aptamer (AP1) and a fumonisin B1 nucleic acid aptamer (AP2) onto the electrode, incubating for 1-10h at 1-10 mu mol/L, v/v, 1/1, and blocking the same with 1 mg/mL QBSA; and (3) finally, dripping a mixed solution of a self-developed ZEN nucleic acid aptamer partial complementary sequence (CP1-Au-Thi) solution of 5'-endmodified gold nanoparticles and thionine and a FB1 nucleic acid aptamer partial complementary sequence (CP2-Au-FC6S) solution of 5'-end modified gold nanoparticles and ferrocenyl hexanethiol, and incubating for 1-10h at 1-10 mu mol/L, v/v, 1/1 to obtain the required aptamer sensor. The method provided by the invention is suitable for the rapid and simultaneous detection of QEN and FB1 in agricultural products such as corn and feed, thereby effectively preventing the agricultural products with over-standard QEN and FB1 from flowing into the market, and the diet safety of people is guaranteed asa result.

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 relates to a chemiluminescence enzyme linked immunosorbent assay kit for detecting Fumonisins. The chemiluminescence kit has the characteristics: a luminescent plate is a detachable 96 aperture opaque white luminescent plate adsorbing a conjugate of Fumonisins and ovalbumin (OVA), and a conjugate coating concentration is 0.25mg /L; a standard substance concentration is 0, 0.1, 0.5, 2.5, 12.25, 61.25 mug / L; a ratio of methanol to PBS is 7:93 (v / v); a chemiluminescence substrate liquid is a mixed solution of luminol and superoxol in a ratio of 1:1; a weak solution is a phosphatic buffer (0.01M, pH 7.4) containing 0.5% BSA; a 10 time concentrated washing liquid is 0.1M phosphatic buffer (containing 0.5% tween-20). The chemiluminescence kit has characteristics of high sensitivity, strong singularity and easy and convenient operation, and can be used to detection of Fumonisins in grain of cereal and products thereof.

The invention discloses a kit for synchronously detecting six fungal toxins. The six toxins are aflatoxin, ochratoxin, patulin, trichothecene mycotoxin, fumonisins and zearalenone; and based on a multiple PCR detection method, the kit comprises a first primer pair, a second primer pair, a third primer pair, a fourth primer pair, a fifth primer pair, a sixth primer pair and an internal standard primer pair which can perform PCR reaction in the same reaction tube. The detection system directly targets a toxin synthesis essential gene and has high specificity; an internal reference gene is amplified synchronously, positive, negative and blank control are set simultaneously, and false negative and false positive are effectively avoided; and multiplication of PCR index guarantees the sensitivity of the method.

The invention discloses stable controllable high-quality Pu'er tea and a fermentation production method thereof. The Pu'er tea is fermented Pu'er tea. In dry base mass, the content of tea polyphenol is 10.0 to 15.0 weight percent, the content of theabrownin is 8.0 to 16.0 weight percent, and the content of a water extract is 30.0 to 45.0 weight percent; in the Pu'er tea, the content of aflatoxin B1 is lower than 0.03mu g/kg, the content of fumonisins B1 is lower than 7mu g/kg, the content of fumonisins B2 is lower than 3mu g/kg, the content of fumonisins B3 is lower than 3mu g/kg, the contentof ochratoxin is lower than 0.3mu g/kg, and the content of vomitoxin is lower than 5mu g/kg. According to the fermentation production method disclosed by the invention, a brand-new fermentation production method of the Pu'er tea is established; the production efficiency is high and the obtained fermentation Pu'er tea has the advantages of high quality, good mouth feel, bright soup color, red-brownleaf bottom and controllable content of important components of materials; in addition, the aflatoxin, the fumonisins, the ochratoxin, the vomitoxin and the like are not detected from the Pu'er tea.

The invention relates to a Fumonisin immunoaffinity column, and a making method and a use thereof. The immunoaffinity column is coupled to an agarose gel carrier by using a protein G, and an anti- Fumonisin antibody is used to couple with the protein G on agarose. The above obtained coupled Fumonisin antibody-protein G-agarose gel carrier composite carrier is filled to an affinity column. The immunoaffinity column is mainly used for purifying Fumonisin in foods, feeds, milk, blood samples and other various samples in order to provide convenience for later stage high performance liquid chromatography (HPLC) detection and fluorescence detection of Fumonisin in the samples.

The invention belongs to the technical field of agricultural biology, and particularly relates to Fumonisin degrading enzyme FumDXA as well as a gene and an application thereof. The Fumonisin degrading enzyme protein FumDXA derived from Xenophilus azovorans is provided, the amino acid sequence of the Fumonisin degrading enzyme protein FumDXA is represented as SEQ ID NO.1, and the encoding gene forencoding the Fumonisin degrading enzyme is also provided. The Fumonisin degrading enzyme FumDXA has activity of degrading Fumonisin B1 and can be applied to agriculture, feed and food industries, sothat harm of Fumonisin to health of animals and humans is reduced.

A multi-analyte column is disclosed. The column may contain at least one unit of resin having ochratoxin specific affinity and, for each unit of resin having ochratoxin specific affinity, the column further contains about 0.95 to 1.05 units of resin containing antibody having specificity for zearalenone, about 1.9 to 2.1 units of resin containing antibody having specificity for aflatoxin and about 2.8 to 3.2 units of resin containing antibody having specificity for fumonisin. One unit of resin is the quantity of resin containing antibody that will bind 50 ng of aflatoxin, 3300 ng of fumonisin, 50 ng of ochratoxin or 1140 ng of zearalenone, respectively.

The invention discloses three specific genes capable of generating fumonisins fusarium microspecies and application thereof and belongs to the field of molecular biology. On the one hand, three specific gene sequences for generation of the fumonisins fusarium microspecies are disclosed; on the other hand, the application of the three specific genes capable of generating the fumonisins fusarium microspecies in detection of rice spike monilinia fiucticola is also disclosed. The three specific genes have the advantages that specific genes of the microspecies are obtained from genomic sequence information of three kinds of fusarium capable of generating fumonisins, detection of pathogenic bacteria is achieved according to specific nucleotide sequences of the microspecies, detection primers have high specificity, and therefore bands with specificity can be amplified from a target strain. The specific genes, the primers and the detection method can be adopted for fast, conveniently and accurately distinguishing three kinds of bacterial strains which can generate fumonisins and trigger rice spike rot.