53 results about "Aflatoxin M" patented technology

The invention discloses a method for degrading aflatoxin, which makes use of gamma rays to irradiate a sample containing the aflatoxin, wherein the gamma rays are generated by a radioactive substance Co, and the irradiation dose is between 0 and 10 kGy not including 0, preferably between 2 and 10 kGy, more preferably between 4 and 10 kGy, particularly preferably between 6 and 10 kGy, and the most preferably 10 kGy. The sample containing the aflatoxin is a farm product containing the aflatoxin or a product obtained by processing the farm product, such as food, feedstuff, and the like. The method is particularly applicable to degrading the aflatoxin B1, not only can kill pathogenic microorganisms but also can degrade biotoxin therein, and does not generate any industrial pollution.

The invention discloses a fluorescent microsphere immunochromatographic test strip for detecting aflatoxin M1. The test strip comprises a sample binding pad, a chromatographic membrane, a water absorption pad and a viscose bottom liner, and the sample binding pad contains a fluorescent microsphere labeled aflatoxin M1 monoclonal antibody; and the chromatographic membrane comprises a detection zone and a quality control zone, the detection zone is coated with an aflatoxin M1 semiantigen-carrier protein conjugate, and the quality control zone is coated with a goat anti-mouse antiantibody. A method for detecting the aflatoxin M1 by using the test strip has the advantages of simplicity, rapidness, wide application range, low cost and easy popularization use.

The invention belongs to the field of biological detection. An AFM1 immunity chromatography test paper strip is characterized in that: the test paper strip comprises a paperboard, an absorbent pad, a detection pad, a colloidal gold pad and a sample pad are sequentially pasted on one side of the paperboard, and adjacent pads overlappingly connect at a joint; the detection pad treats a cellulose nitrate membrane as a base pad, a quality control wire and a detection wire are arranged on the cellulose nitrate membrane in a top-down manner, the detection wire is coated with an AFM1-bovine serum albumin (AFM1-BSA) conjugate, and the quality control wire is coated with a rabbit-anti-mouse polyclonal antibody; and the colloidal gold pad is transversely sprayed with a nanogold labeled AFM1 monoclonal antibody which is generated by a hybridoma cell strain 2C9 with the accession number of CCTCCNO.C201018. The test paper strip which is used for AFM1 detection has the characteristics of rapid detection, simple operation and high sensitivity.

The invention belongs to the field of food safety monitoring, and discloses an immunochromatographic colloidal gold test strip for detecting aflatoxin M1. The test strip includes a sample pad, a nitrocellulose membrane (NC), an absorbent pad and a PVC backing. The nitrocellulose membrane is adhered to the on PVC backing; and the sample pad and the absorbent pad are adhered to the ends of the nitrocellulose membrane. The colloidal gold strip uses a one-step indirect competitive immunochromatographic technology for rapid detection of whether the aflatoxin M1 residue in milk meets the EU limit standard (less than 0.05ng / mL) and the limit standards of China (the United States and other countries) (less than 0.5n / mL), and has sensitivity up to 0.05ng / mL.

The invention relates to a strain of tetragenococcus halophilus which can effectively remove aflatoxin B1 (AFB1) in a high-salt environment. First of all, an activated strain of tetragenococcus halophilus is cultivated in an MRS fluid medium until entering the early logarithmic phase, and then applied respectively to a high-salt liquid material system added with pure AFB1 toxin and a high-salt semisolid material system contaminated by AFB1 so as to remove AFB1; finally, the strain of tetragenococcus halophilus is used in a kind of thick broad-bean sauce contaminated by AFB1. In a simulated factory production environment, the removal rate of AFB1 by the tetragenococcus halophilus of the invention can reach 41.76%. The invention is applicable to the traditional brewage industry, especially in the flavouring industry with high-salt environments, and has good economic benefit and practical application value.

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 chemiluminescence enzyme-linked immune detection kit of aflatoxin M1 and a using method, belonging to the technical field of chemiluminescence enzyme-linked immune detection. The detection kit comprises a chemiluminescence enzyme label plate coated with aflatoxin M1 antigen, an aflatoxin M1 standard, an aflatoxin M1 antibody, an enzyme-labeled antibody, a chemiluminescent solution A and a chemiluminescent solution b. The using method of the kit comprises the following steps of: (1) pre-processing the samples to be detected; (2) orderly adding the aflatoxin M1 standard solutions or samples and the aflatoxin M1 antibody, adding the enzyme-labeled antibody after a competing reaction, finally adding the chemiluminescent solutions for quantitative detection of the aflatoxin M1 by a chemiluminescence immunoassay analyzer; and (3) processing and analyzing the result. The chemiluminescence enzyme-linked immune detection kit of aflatoxin M1 disclosed by the invention has advantages of high sensitivity, good stability and important practical application and development values, and is suitable for screening a large number of samples.

The invention discloses a test strip and a method for detecting aflatoxin M1. The test strip comprises test paper and a micro-pore reagent, wherein a colloidal gold marked aflatoxin M1 monoclonal antibody is formed in the micro-pore reagent through freeze-drying; the test paper is composed of a sample absorption pad, a reaction film, a water absorption pad, a protection film and a bottom plate, which are sequentially connected; the reaction film comprises a detection area and a quality control area, wherein the detection area is coated with an aflatoxin M1 semi-antigen-carrier protein conjugate, and the quality control area is coated with an anti-antibody. The method for detecting the aflatoxin M1 by the test strip disclosed by the invention is simple, fast, direct and accurate; furthermore, the test strip has wide application scope and low cast and is suitable for popularizing and using.

The invention relates to a detection gel column of aflatoxin M1 and a detection method of the aflatoxin M1. The detection gel column of the aflatoxin M1 comprises a hollow column as well as a detection layer and a quality control layer that are filled into the hollow column. When in detection, a to-be-detected sample and an aflatoxin M1 enzyme-labelled antigen marked by a chemiluminiscent label are added into the detection gel column of the aflatoxin M1 and flow through the detection layer and the quality control layer. Both the to-be-detected sample and the aflatoxin M1 enzyme-labelled antigen marked by the chemiluminiscent label contend an aflatoxin M1 monoclonal antibody, and whether the to-be-detected sample contains the aflatoxin M1 is judged according to the color of the detection layer and the color of the quality control layer. The detection gel column of the aflatoxin M1 is carried conveniently, can be used for detecting whether the to-be-detected sample contains the aflatoxinM1 rapidly and sensitively and is suitable for on-site rapid detection of a large number of samples.

The invention discloses a method and an enzyme linked immunosorbent assay kit for detecting the content of aflatoxin M1 in a sample. The operation steps can be decreased by adopting direct competitive ELISA detection mode through adopting high-specificity and high-affinity antibodies, and the detection sensitivity and accuracy can be enhanced; compared with the antibody coating, the coating of an ELISA plate is carried out by adopting coating antigen, the better coating effect and long storage time can be more favorably achieved, and further the detection precision and stability of the kit can be enhanced; in addition, according to the kit, marking is carried out by adopting ELISA plate labelled antibody technology through adopting an improved periodate oxidization method, enzyme is directly labelled on an aflatoxin M1 specificity antibody, the two most important reactants namely aflatoxin M1 specificity antibody and enzyme are combined into a whole, so that the labelling efficiency is improved, the usage amounts of the enzyme and the antibody are saved, the good activities of the labelled enzyme and the antibody are guaranteed, the antibody is unnecessarily arranged in the kit, and the cost of the kit is greatly lowered.

An aflatoxin M1 nanobody, an immunosorbent and an immunoaffinity column. The aflatoxin M1 nanobody 2014AFM-G2 has the amino acid sequence of SEQ ID NO:7, is encoded by the nucleic acid sequence of SEQ IDNO:8, has a 50% inhibiting concentration IC₅₀ to aflatoxin M1 of 0.208 ng/mL, and has cross reaction rates with aflatoxins B1, B2, G1, and G2 of 9.43%, 5.93%, 4.87% and 6.17%, respectively. The immunosorbent includes a solid phase carrier and aflatoxin M1 nanobody 2014AFM-G2 coupled with the solid phase carrier. The immunoaffinity column is loaded with the aflatoxin M1 nanobody immunosorbent. It can be used for purifying and concentrating an extracting solution of a sample before loading to a machine for detection and the immunoaffinity column can be used repeatedly for many times.

The invention discloses an aflatoxin M1 gold label quick detection card and a preparation method and an application thereof, belongs to the field of immunology, and relates to a toxin detection technology. The detection card comprises a detection strip and a plastic card casing, wherein the detection strip is supported by a PVC (polyvinyl chloride) bottom lining and consists of a sample pad, a gold label antibody combination pad, an enveloping membrane and a water absorption pad which are sequentially connected; the gold label antibody combination pad is made of glass fibers, and envelops an aflatoxin M1 monoclonal antibody combined with colloidal gold particles; the enveloping membrane is a cellulose nitrate membrane and envelops a detection line (T line) containing recessive aflatoxin M1 protein conjugates and a control line (C line) containing a goat-anti-mouse monoclonal antibody; the sample pad is made of glass fibers processed by a buffering system. The detection card, based on the colloidal gold immunochromatograohic assay technology, is simple to operate, convenient to carry, and quick and accurate in result determination, requires only 40-50 minutes for detection, and is suitable for on-site supervision and qualitative screening of a great number of samples.

The invention relates to a magnetic immuno-chromatographic kit for detecting aflatoxin M1 (AFM1). The magnetic immuno-chromatographic kit for detecting AFM1 comprises at least one test strip and the solution of a superparamagnetic composite particle labeled AFM1 antibody, wherein the test strip is assembled by adhering a reaction pad, a sample pad 1, a sample pad 2 and an absorbent pad to a base board in sequence in a mutual staggered way and covering the pads with a transparent plastic film, the reaction pad is pre-coated with an AFM1 protein conjugate detection line and an AFM1 antibody combined anti-antibody quantity control line, and the superparamagnetic composite particle labeled AFM1 antibody is a polymer which is formed by combining AFM1 antibody with superparamagnetic composite particles by peptide bonds covalently. The invention further relates to a preparation method of the magnetic immuno-chromatographic kit and application of the magnetic immuno-chromatographic kit to detection of AFM1 in dairy products. The kit is used for immuno-chromatographic detection by taking magnetic particles as labels and has the advantages of high detection speed, convenience in operation, wide linear range and high sensitivity.

The invention provides a method for detecting aflatoxin M1. The method is based on a direct competitive ELISA (enzyme-linked immunosorbent assay) technique, firstly, after monoclonal antibodies are coated, a to-be-detected sample and aflatoxin M1 labeled with catalase C100 are added, aflatoxin M1 in the sample and the aflatoxin M1 labeled with catalase C100 are competitively combined with the monoclonal antibodies fixed on an ELISA plate, fluorescence quenching of cadmium telluride quantum dots modified with mercaptopropionic acid is reduced through decomposition of hydrogen peroxide under the catalytic function of catalase, and the content of aflatoxin M1 in the sample is judged according to the fluorescence intensity. According to the method, new catalase is introduced innovatively, and the reaction precision is improved while the cost is reduced; meanwhile, a more sensitive novel fluorogenic substrate, namely, the cadmium telluride quantum dots modified with mercaptopropionic acid, is adopted, and the lighting sensitivity is remarkably improved by comparison with traditional TMB (tetramethylbenzidine) substrates.

Aflatoxin M1 nanobody 2014AFM-G2 has the amino acid sequence of SEQ ID NO:7, and is encoded by the gene sequence of SEQ ID NO:8. The aflatoxin M1 nanobody 2014AFM-G2 obtained via screening has the properties of tolerance to organic reagents, tolerance to high temperature, tolerance to acids and bases and the like, and good stability. The aflatoxin M1 nanobody 2014AFM-G2 has 50% inhibiting concentration IC₅₀ to aflatoxin M1 of 0.208 ng/mL, and has cross reaction rates with aflatoxin B1, B2, G1, G2 are 9.43%, 5.93%, 4.87% and 6.17%, respectively.

The invention relates to an aflatoxin B1 detection kit, and belongs to the technical field of enzyme-linked immunosorbent assay (ELISA), wherein the aflatoxin B1 detection kit is used for detecting aflatoxin B1 (short for AFB1) content in grains, feeds and food. The aflatoxin B1 detection kit comprises a box body, a 24-cell AFB1 coated reaction plate inside the box, a reaction cover plate, 13 bottles of reagents, concave bottle positions for placing the reagents, an ice bag, a frame, an instruction manual, a packet of water absorption paper, two disposable droppers and a quality detection report. The aflatoxin B1 detection kit is characterized in that the coated reaction plate adopts a 24-well reagent plate as a solid phase carrier, and the 13 bottles of the reagents comprise 6 bottles of AFB1 standard substance solutions, enzyme labeled antigen, an enzyme labeled antigen dilution buffer solution, a concentration washing solution, a sample dilution solution, a substrate solution, a coloration solution and a termination solution, and the number of the concave bottle positions is 16. The kit has characteristics of simple structure, easy use, low price, and high sensitivity, wherein the sensitivity can be more than 0.l ng/ml.

The invention discloses an aptamer for detecting aflatoxin M1, a sensor, a kit and application thereof. The aptamer for detecting aflatoxin M1 has the nucleotide as shown in SEQ ID No:1. The invention further discloses a sensor for preparing the nucleotide aptamer and a detection kit containing the sensor solution. Signal amplification is carried out by using PCR technology as signal transduction, the detection limit is greatly reduced and reaches 0.03ng/L. The nucleotide aptamer is used as an identification unit, the sensor prepared from the nucleotide aptamer is used for detecting the content of aflatoxin M1 in a sample and has high sensitivity and wide linear range when detecting the aflatoxin M1, and accurate quantitative detection can be realized.

The invention discloses a method for detecting aflatoxin M1. The invention provides application of a kit to detection on the aflatoxin M1. The kit contains a fluorescently-labeled anti-aflatoxin M1 antibody, a superparamagnetic particle labeled aflatoxin M1 coupling protein antigen and a micro-fluidic chip detection card; the micro-fluidic chip detection card comprises a bottom plate and a micro-fluidic chip; the micro-fluidic chip is provided with a detection passage; the detection passage is formed by communicating a bottom passage with a sampling passage and a liquid suction passage which are respectively positioned at both ends of the bottom passage; openings of the sampling passage and the liquid suction passage respectively are a sampling hole and a liquid suction hole; the bottom passage is fixedly provided with a filter element; and the filter element is provided with a plurality of through holes for enabling sampling liquid to flow through. The detection kit and the detectionmethod, which are provided by the invention, are used for detecting the aflatoxin M1, and are high in detection sensitivity, short in detection time, low in detection cost and easy to operate and popularize.

The invention discloses a chemiluminescent enzyme immunoassay (CLEIA) detection kit for aflatoxin M1. The kit comprises a kit body, a chemiluminescent plate arranged in the kit body and a reagent arranged in the kit body. The kit is characterized in that each hole in the chemiluminescent plate is coated with an anti-aflatoxin M1 antibody and the reagent comprises an enzyme-labeled aflatoxin M1 antigen concentrated solution, an enzyme-labeled aflatoxin M1 antigen diluent solution, aflatoxin M1 series standard solutions, a chemiluminescent substrate liquid A, a chemiluminescent substrate liquid B, a concentrated washing solution and a concentrated complex solution. The chemiluminescent enzyme immunoassay detection kit has the characteristics of high sensitivity, simplicity, rapidness and high accuracy. Compared with traditional ELISA methods, the kit provided by the invention enables operation time to be greatly reduced. The chemiluminescent enzyme immunoassay detection kit can be used for detecting aflatoxin M1 residue in milk and milk powder.

The invention discloses an aflatoxin M1 fluorescent sensitizer. The aflatoxin M1 fluorescent sensitizer is composed of methyl-beta-cyclodextrin and a Hg<2+> containing solution, wherein the mole ratio of methyl-beta-cyclodextrin to AFM1 is (5.8*10<5>):1; the mole ratio of Hg<2+> to AFM1 is (6.6*10<4>):1. The Hg<2+>-M-beta-CD fluorescent sensitizer has a remarkable reinforcing effect on aflatoxin M1 fluorescence and the reaction is quick. A related coefficient R of an AFM1 to-be-detected solution system can reach 0.999 or above in the concentration range of 0.01-2 [mu]g/L and the detection limit is 0.0026 [mu]g/L. The quick, sensitive and economic fluorescence reinforcing agent is expected to be directly applied to detection of AFM1 in dairy products and a new idea and a new approach are provided for rapid detection of AFM1.

The invention belongs to the field of milk and milk product quality safety detection, and particularly discloses a special purification column for an aflatoxin M group and an application. The specialpurification column comprises a column pipe; a lower sieve plate, a filler and an upper sieve plate are sequentially arranged in the column pipe from the liquid outlet end of the column pipe to the liquid inlet end of the column pipe; and the filler is a mixture of graphitized carbon and nano ferroferric oxide. The invention also discloses an application of the special purification column to the purification and detection of the aflatoxin M group in milk and milk products. The special purification column for the aflatoxin M group is simple and convenient to manufacture and low in cost, is applied to the detection of the milk and milk products, can simplify operation and shorten detection time, can specifically adsorb AFM1 and AFM2, and has excellent adsorption effect; impurities in sampleintroduction are greatly reduced after being purified by the special purification column; and the special purification column has extremely strong practical popularization significance.

The invention discloses a method for treating aflatoxin B1 in stored corn by using chlorine dioxide, which belongs to the technical field of food engineering. The method comprises the steps that cornis flatly laid in a closed storage chamber, and the corn is in a flowing type; chlorine dioxide gas with a concentration of 60-100mg/L is introduced into the closed storage chamber, and fumigating iscarried out for 12-24 hours in the closed storage chamber; and finally, ventilation is conducted, and the corn storage conditions in the closed storage chamber are set as follows: under the standard atmospheric pressure, the humidity is 60-75%, and the temperature is 20 DEG C or below. According to the method, the chlorine dioxide gas is used as a degradation substance of the aflatoxin B1, the aflatoxin B1 in the stored corn is treated under the conditions of specific chlorine dioxide gas concentration, treatment time and the like, and the degradation rate of the aflatoxin B1 can reach 78%; and meanwhile, the processing method is suitable for large-scale corn storage.

The invention provides an anti-aflatoxin M1 monoclonal antibody immunoadsorbent, an immunoaffinity column and a preparation method thereof. The anti-AFM1 immunoadsorbent comprises a solid phase carrier and an anti-aflatoxin M1 monoclonal antibody conjugated to the solid phase carrier. The solid phase carrier is an epoxy group-containing activated resin. The anti-aflatoxin M1 monoclonal antibody has an IgG2b type heavy chain and a gamma-type light chain. The front 15 amino acid sequence of the amino group end of the light chain is GVTQESALTTSPGGT. The front 15 amino acid sequence of the amino group end of the heavy chain is EVILVESGGGLVKPG. The molecular weight is 150KD. A coupling ratio of the anti-aflatoxin M1 monoclonal antibody and activated resin is 93.4%. The anti-aflatoxin M1 monoclonal antibody immunoadsorbent has a milk aflatoxin M1 adsorption capacity of 100 microgram per milligram antibody. The immunoadsorbent can remove aflatoxin M1 in fresh milk processing without loss of fresh milk nutrition.

The invention discloses a solid-phase extraction column and a solid-phase extraction column filling material production method and a method for detecting aflatoxin by using the solid-phase extractioncolumn thereof, which belong to the field of food detection. The solid-phase extraction column comprises a solid-phase extraction column sleeve, an injection sample-introduction port, and a liquid outlet, a cover is arranged at the solid-phase extraction column sleeve, the injection sample-introduction port is arranged at the cover, an upper glass fiber baffle plate and a lower glass fiber baffleplate are arranged in an inner chamber of the solid-phase extraction column sleeve, and a solid-phase extraction mixed filling material is placed between the upper glass fiber baffle plate and the lower glass fiber baffle plate. The method for producing the solid-phase extraction column filling material comprises a carrier synthesis step and an ionic liquid immobilization step. The processing timeof the superfast solid-phase extraction column is short, the detection cost is low, and the method has good reappearance and stability. The rapid extraction method completely uses a high-efficiency extraction technology, which is stable and fast. The solid-phase extraction column is used for detecting the content of aflatoxin in milk and a dairy product.

The invention discloses a time resolution fluorescence immunoassay kit for detecting aflatoxin M1. The time resolution fluorescence immunoassay kit for detecting aflatoxin M1 consists of a porous coating plate, a buffer solution, an aflatoxin M1 standard substance, an aflatoxin M1 antibody freeze-drying product, a europium-labeled goat anti-rat antibody, a scrubbing solution and an enhancing solution. A detection method of the time resolution fluorescence immunoassay kit for detecting aflatoxin M1 comprises the following steps: (1) preparation of an immunogen; (2) preparation of a coating source; (3) preparation of a monoclonal antibody; and (4) pretreatment and detection of a sample. The time resolution fluorescence immunoassay kit is short in detection time, high in average recovery rate, simple in sample pretreatment, wide in application, low in detection cost, high in detection specificity, small in intra-batch and batch-to-batch difference, high in sensitivity and simple and quickin operation, is capable of field operation detection, and is particularly applicable for the detection of large-scale samples.