91 results about "Fumonisin B1" 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 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 relates to a reagent box for quantificationally detecting the content of fumonisin B1 in food by adopting an enzymoimmunoassay technology and the detecting method thereof. The reagent box includes the steps that a fumonisin B1-KLH antigen is prepared; a polyclonal fumonisin B1 antibody is prepared; an enzyme labeling antigen is prepared; a coated plate is prepared, and a reagent is prepared. The invention mainly adopts the enzymoimmunoassay technology to detect the fumonisin B1, can realize the aim of quantificationally detecting the content of fumonisin B1 in food and alimentary crops; and the reagent box has simple structure, convenient usage, cheap price and high sensitivity; the detecting method has strong specificity, high sensitivity, good veracity, and the sensitivity can reach gamma/kg grade.

The present invention relates to an immunoadsorbent and a composite affinity column for purification of fumonisin B1, aflatoxin, ochratoxin A and zearalenone. The immunoadsorbent comprises a solid carrier and fumonisin B1, aflatoxin, ochratoxin A and zearalenone monoclonal antibodies coupled with the solid carrier, the fumonisin B1 monoclonal antibody is secreted from hybridoma cell line Fm7A11 with the preservation number of CCTCCNO.C201636, the hybridoma cell line Fm7A11 is preserved in CCTCC in March 29, 2016 in Wuhan University Wuhan China. The composite affinity column is loaded with the immunoadsorbent with the fumonisin B1, aflatoxin, ochratoxin A and zearalenone monoclonal antibodies, and is used for purification of the fumonisin B1, aflatoxin, ochratoxin A and zearalenone. The composite affinity column can be used for purification and detection of fumonisin B1, aflatoxin, ochratoxin A and zearalenone toxin samples.

The invention discloses a time-resolved fluorescence immunoassay kit for detecting fumonisins B1 and a detection method thereof, belonging to the technical field of time-resolved fluoroimmunoassay (TRFIA) for detecting the content of FB1 in grain, fodder and food. The prepared kit adopts the TRFIA to detect the FB1. A micropore plate is coated with FB1-BSA, is added with FB1 standard or sample, and is added with FB1 monoclonal antibody. Dissociative FB1 and the FB1-BSA on the micropore plate compete for the FB1 monoclonal antibody, unconnected FB1 monoclonal antibody is removed by means of washing, Eu3+ -sheep anti-mouse antibody is added, and the unconnected Eu3+ -sheep anti-mouse antibody is removed by means of washing after labeled immune reaction. After adding strengthening liquid, fluorescence intensity (cps) is detected by a time-resolved fluoroimmunoassay apparatus, wherein the fluorescence intensity is in inverse proportion to the concentration of the FB1, and the content in the FB1 can be ensured by contrasting a standard curve. The kit has simple structure, convenient use, low price and high sensitivity more than 0.05ng/mL.

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.

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.

The invention provides an anti-aflatoxin general type monoclonal antibody hybridoma cell line and application thereof, belonging to the field of food safety immunodetection. The cell line 1G6 provided by the invention is collected in the China General Microbiological Culture Collection Center (CGMCC) with a collection number of CGMCC No.7210. After an aflatoxin complete antigen is uniformly mixed with equivalent Quick Antibody immunologic adjuvant TM, an obtained mixture is injected to immunize BALB/c mice through leg muscle. B1-KLH (fumonisin B1-Keyhole Limpet Hemocyanin) is adopted for the first time of immunization, M1-KLH (fumonisin M1-Keyhole Limpet Hemocyanin) is adopted for the second time of immunization, and an M1 complete antigen (without containing adjuvant) is adopted for the last time of impaction immunization. Splenocytes of immunized mice are fused with myeloma cells through a PEG (polyethylene glycol) method, and the hybridoma cell line 1G6 is obtained through indirect ELISA (Enzyme-Linked Immuno Sorbent Assay) and indirect competence ELISA screening and three times of subcloning. A monoclonal antibody secreted from the cell line has better affinity and detection sensitivity for aflatoxins B1, B2, G1, G2 and M1, and can be used for immunodetection of the total quantity of the aflatoxins in the food safety.

The invention relates to a quantitative detection method of fumonisins B1, comprising the following steps of: 1, preparing conjugates FB1-KLH and FB1-OVA; 2, preparing anti-FB1 monoclonal antibody; 3, preparing colloid gold, labeling the anti-FB1 monoclonal antibody and spraying the labeled monoclonal antibody onto a gold labeled gasket; 4, spotting on a nitrocellulose membrane; 5, assembling to form strips; and 6, drawing a standard curve of FB1 concentration and color value and dragging the color value of a sample to be tested into the standard curve to obtain the FB1 content in the sample to be tested. The invention has advantages of single detection object and strong pertinence, high accuracy, fast detection speed and short required time. The method provided by the invention can be used for detection without trained professionals. The method satisfies the demand of rapid and correct judgment of FB1 content in food storage and sale institution, exit-entry and custom inspection departments, and is convenient for popularization and application at grass-root level.

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 discloses a preparation method of a golden flower nanoparticle immune chromatography test strip capable of simultaneously detecting deoxynivalenol and fumonisins B1 in grains. The method comprises the following steps: (1) preparing a DON monoclonal antibody-golden flower nanoparticle marker and a FB1-monoclonal antibody-golden flower nanoparticle marker; (2) treating a gold-marked conjugate release pad; (3) enveloping a nitrocellulose membrane; and (4) preparing the test strip. The immune chromatography test strip is high in specificity and sensitivity, visual and intuitive in detection result, simple, convenient and fast to operate, and easy to popularize and apply within a large range, and is widely suitable for the requirements such as grain purchase site inspection, food safety inspection and customs quarantine control.

The invention relates to an aptamer combined with high specificity and high affinity of Fumonisins B1 and applications thereof. The aptamer can be used for detection and analysis and separation and concentration of the Fumonisins B1, and has the characteristics of high specificity, high affinity, uniform quality, high stability, short development cycle, low production cost and convenience in use. The invention also relates to sequences derived from aptamer sequences, which comprise modified sequences.

The invention relates to the technical field of analysis and detection, and in particular, relates to a detection kit for multiple fungaltoxins, and a preparation method, a detection method and application of the detection kit. The kit comprises up-conversion nanoparticles respectively coupled with antigens of fungaltoxins to be detected, magnetic nanoparticles respectively coupled with antibodies of the fungaltoxins to be detected, and a standard substance solution of the fungaltoxins, and the fungaltoxins to be detected at least comprise zearalenone and fumonisin B1. The kit and the method have the advantages of high sensitivity, good specificity and the like, have important practical significance on ultra-sensitive detection of multiple fungaltoxins, and have good guiding significance on realization of a field rapid and ultra-sensitive detection technology.

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.

The invention provides a reagent kit for detecting Fumonisin B1 of genetically engineered single-chain antibody and method thereof; in the reagent kit of the invention, the reagent comprises: sample extracting solution, a standard reagent, an enzyme labeled antigen reagent, washing solution, a BSA (bovine serum albumin) regent, a chromogenic substrate and stopping solution; the content of FB1 toxins in the sample is calculated by the following steps of sample processing, regent balancing, washing, adding sampling, washing, color developing and stopping. The Fumonisin B1 single-chain antibody adopted by the reagent kit provided by the invention can be effectively expressed in colon bacillus, and can be produced in a large scale; the producing process is simple, convenient and easy; the time, labor and money are saved. Using the reagent kit in the invention for detecting the Fumonisin B1, whether the sample is polluted by the Fumonisin B1 or not can be determined, and the content of the Fumonisin B1 is calculated in 1.5-2h. The reagent kit is convenient and fast in use, and low in cost.

The invention relates to a method for simultaneously detecting fumonisins B1 and B2 in different matrix traditional Chinese medicines, namely an on-line derivation high-performance liquid phase fluorescent detection method. The method comprises the steps of extracting, purifying and detecting a sample and particularly comprises the following steps of: adding sodium chloride in the sample; performing oscillating extraction by using a horizontal table according to the volume ratio of methanol to water of 4:1; filtering; diluting a certain amount of filtrate by adding water; filtering by using a microfiber filtration membrane; purifying by using an immunoaffinity column; drying by using nitrogen; dissolving by using methanol; and sampling. A derivation agent is prepared by the following steps of: dissolving a proper amount of o-phthaldialdehyde by using methanol; adding into a proper amount of sodium tetraborate solution; adding 2-mercaptoethanol; mixing uniformly; and passing through a microfiltration membrane. The measurement method of the sample comprises the following steps of: separating by using C18 reversed phase chromatographic column; performing gradient elution by using methanol and sodium dihydrogen phosphate as mobile phase; reacting with the derivation agent; detecting by using a fluorescence detector; and confirming the positive sample by using high-performance liquid chromatography-mass spectrum.

The invention discloses an immunoassay kit and a special antibody for detecting fumonisins. A single-chain antibody is prepared by sequentially connecting a heavy chain variable region, short peptide connecting the heavy chain variable region and a light chain variable region, and the light chain variable region, wherein the amino acid sequence of the heavy chain variable region is represented by 1st to 118th amino acid residues of a sequence 1 from an N end; and the amino acid sequence of the light chain variable region is represented by 134th to 246th amino acid residues of the sequence 1 from the N end. The affinity constant of the antibody is 1.73*10<9>L/ mol, and the half abatement (IC50) is 0.15 ng/mL. In the invention, antibody sources with high valence and high specificity are provided for establishing a method for detecting fumonisins residues in food. The kit has the advantages of high sensitivity, high accuracy, high precision, high specificity for fumonisins B1 and low cost. Therefore, the antibody, the kit and the detection method play an important role in detecting the fumonisins B1.

The invention relates to a method for simultaneous quantitative detection of zearalenone and fumonisin B1. The method comprises the steps of: 1. preparation of conjugates ZEN-BSA and ZEN-OVA, FB1-KLH and FB1-OVA; 2. preparing an anti-ZEN monoclonal antibody and an anti-FB1monoclonal antibody; 3. preparing colloidal gold, labelling the monoclonal antibodies, and spraying the labelled monoclonal antibodies to a gold labeled pad; 4. conducting sample application on a cellulose nitrate membrane; 5. carrying out assembling to obtain a test strip; and 6. respectively drawing standard curves of the concentrations and color values of ZEN and FB1, bringing the color value of a sample to be tested into the standard curves so as to obtain the content of ZEN and FB1 in the sample to be tested. The method of the invention has the advantages of high detection accuracy, fast detection speed, and short detection time, and can be used without training, thus meeting the demands for rapid and correct judging of ZEN and FB1 content of grain storage and sales organizations, as well as immigration, customs and other inspection departments.

The invention provides a sensitivity detection method for fumonisin B1. The method includes: utilizing fluorescent microspheres embedded with quantum dots to replace conventional enzyme carriers to be coupled with fumonisin B1; using a coupling product as a competition antigen to execute direct competition ELISA (enzyme-linked immune sorbent assay). In a technical path, proper quantum dots are selected according to luminescent characteristics, a quantum dot embedding method based on fluorescent microsphere technology is further designed, and on this basis, quantum dot fluorescent microspheres are subjected to BSA embedding and then coupled with fumonisin B1 to obtain the competition antigen with better performance. In the technical scheme, a lot of quantum dots are embedded in the fluorescent microspheres through a high polymer carrier, so that the fluorescent microspheres have higher illumination intensity, and detection sensitivity can be improved effectively; the fluorescent microspheres have large grain size, so that over-high affinity between the competition antigen and an embedding antibody can be lowered to a certain degree, and detection sensitivity is improved.

The invention discloses a composite nano material modified screen print electrode and a method for detecting Fumonisin B1. A working electrode of the screen print electrode is coated with a multi-walled carbon nano tube-colloid gold-chitosan mixture film. The preparation process includes the following steps; activating the multi-walled carbon nano tube; preparing the colloid gold; uniformly mixing the multi-walled carbon nano tube and the colloid gold through the chitosan; coating the mixture on the surface of the working electrode; and completing the preparation after the working electrode is dried at 37 DEG C and the film is formed. The method for detecting Fumonisin B1 provided by the invention is realized in the way that if FB1 exists in a to-be-detected sample, the FB1 competes with FB1 antibodies in the combination with FB1-OVA, so that the antibody amount combined with the FB1-OVA on the electrode is reduced, the amount of HRP-goat-anti-mouse second antibodies on the electrode is reduced, and the current variation value delta I is influenced; and then the content of FB1 in the to-be-detected sample of grains can be judged as per the standard curve drawn through an FB1 standard product. According to the invention, the detection object is single, the pertinence is strong, the accuracy is high, and the sensitivity is strong.