38 results about "Aflatoxicol M1" patented technology

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 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 discloses a method for simultaneously determining aflatoxins and flavoring agents in vegetable oil based on liquid-liquid extraction-liquid chromatography-tandem mass spectrometry. The method can simultaneously determine aflatoxin B1, aflatoxin B2, aflatoxin G1, aflatoxin G2, aflatoxin M1, aflatoxin M2, vanillin, methyl vanillin and ethyl vanillin in the vegetable oil. The method comprises the following steps: carrying out extraction on a sample through using an acetonitrile solvent, carrying out ultrasonically assisted extraction, purifying the obtained extraction solution by n-hexane, performing analysis under gradient conditions, carrying out qualitative analysis through comparing target analytes in retention time and ion pair information in a multi-reaction monitoring manner, and carrying out external standard method quantification. The method has the advantages of small amounts of the sample and reagents, fast analysis speed, high sensitivity, good reappearance and realization of simultaneous detection of aflatoxins and commonly used flavoring agents, and provides a new detection method for evaluating the quality and the safety of vegetable oil 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 discloses a chemiluminescence detection kit for aflatoxins M1 and a preparation method of the chemiluminescence detection kit. The chemiluminescence detection kit comprises an acridiniumester marker, magnetic particles coupled with an antigen or antibody, a calibrator solution, a cleanout fluid, a chemiluminescence pre-excitation liquid A and a chemiluminescence excitation liquid B.According to the chemiluminescence detection kit for aflatoxins M1, the magnetic separation chemiluminescence technology is taken as a detection means, and the acridinium ester marking technology isused together. The chemiluminescence detection kit for aflatoxins M1 is simple and convenient to operate, mild in reaction condition and stable in lighting value and is less influenced by external conditions. Compared with the prior art, the chemiluminescence detection kit for aflatoxins M1 has the advantages of being high in sensitivity, quick and convenient in detection, high in accuracy, good in repeatability and strong in specificity.

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 the field of aflatoxin detection and analysis, and in particular relates to a method for detecting aflatoxins in a vegetable protein beverage. The method comprises the following steps: taking a vegetable protein beverage sample, adding a methanol water solution containing trichloroacetic acid to extract a purification solution, ultrasonically extracting, centrifuging, taking a clear solution for dilution, purifying by an immunoaffinity column, and using high performance liquid chromatography for detecting the content of the aflatoxins. The method can simultaneously detect aflatoxins M1, B1, B2, G1 and G2 in the vegetable protein beverage, and is accurate, rapid, safe and strong in anti-interference ability. The detection limit is 0.05 mug / kg, the specificity is high, and the repetition is good, and the recovery rate is 96.00 to 104.80%. The method can meet the testing and supervision needs of production enterprises and governments, and is of great significanceto ensuring consumer safety.

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 an aflatoxin M1 detection method and a kit. The method includes: hybridizing aflatoxin M1 aptamer (Apt) and single-chain signal probe DNA (ssDNA) to form a hybrid chain Apt-ssCNA; putting the hybrid chain Apt-ssCNA into a to-be-tested sample, when AFM1 exists, allowing reaction between the AFM1 and an Apt segment of the hybrid chain Apt-ssCNA to generate an Apt-AFM1 compound, and releasing the single-chain signal probe DNA (ssDNA); subjecting the hybrid chain Apt-ssCNA to DNA amplification to generate double-stranded DNA, and quickly hydrolyzing the double-stranded DNA into mononucleotides under selective catalytic action of exonuclease, wherein the single-chain signal probe DNA are retained and not hydrolyzed; under ssDNA induction, allowing silver ion reduction to generate near infrared fluorescent silver nanoclusters; detecting system fluorescence intensity, and detecting AFM1 content of the to-be-tested sample according to a relation between the fluorescence intensity and the AFM1 content. The aflatoxin M1 detection method has the advantages of high sensitivity, simplicity in operation and low cost.

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 provides a high-flux enzyme linked immunosorbent assay kit for aflatoxin M1 in milk and a detection method of the kit. The kit comprises a closing solution, an enzyme-labeled antibody concentrated solution, an enzyme label dilution solution, a ELISA plate coated with coating antigens and a substrate developing liquid, wherein the closing solution contains fetal calf serum, skim milkpowder and a protein stabilizer, each 1000mL of the closing solution contains 32.0mL-48.0mL of the fetal calf serum, 7.2g-10.8g of the skim milk powder and 2.4g-3.6g of the protein stabilizer; and theenzyme label dilution solution contains enzyme-labeled antibodies, fetal calf serum and Proclin300, and each 1000mL of the enzyme label dilution solution contains 50mL-100mL of the enzyme-labeled antibodies and 100mu L-400mu L of Proclin300.

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.

The invention discloses a method for detecting aflatoxin M1 in cheese. Extraction, purification and enrichment methods for aflatoxin in cheese are studied, and the aflatoxin M1 in the cheese is detected by combining extraction via hydrophilic-lipophilic balance column with high performance liquid chromatography. The hydrophilic-lipophilic balance column is used to extract the aflatoxin M1. Compared with a aflatoxin M1 test method in the present standard, an OASIS HLB solid-phase extraction purification column has a better separation and purification effect for the aflatoxin M1; the aflatoxin M1 in the cheese can be rapidly detected within 8 min, the average recovery rate is high, the relative standard deviation (RSD) is 1.4%-5.7%, and the lowest detection limit is 0.037 microgram / kg. Compared with the analysis time (100 min) of a second method in the current national standard GB5413.37-2010, detection time is shortened by 40min. The detection method is characterized by short analysis time, simple and rapid operation, low detection cost and accurate detection.

The invention discloses a novel method for quickly detecting aflatoxin M1. The method is used for detecting aflatoxin M1 mainly on the basis of non-labeled porphyrin fluorescent dyes. According to the method, after aflatoxin M1 and porphyrin fluorescent dyes are mixed, the fluorescence intensity of the dyes increases, and the fluorescence intensity is detected to implement quick and quantitative detection on the aflatoxin M1. The method disclosed by the invention is simple, has the advantages of high specificity and high detection speed, and can satisfy the quick and accurate field detection requirements.

The invention discloses a time and fluorescence resolution fluorescence immunoassay kit for detecting aflatoxins M1. The time resolution fluorescence immunoassay kit for detecting the aflatoxins M1 iscomposed of a porous coating plate, buffer liquid, an aflatoxins M1 standard substance, an aflatoxins M1 antibody freeze-drying substance, a europium-labeled goat anti-mouse antibody, a cleaning solution and an enhancement solution. A detecting method of the time and fluorescence resolution fluorescence immunoassay kit for detecting the aflatoxins M1 comprises the following steps that (1) preparation of immunogen; (2) preparation of coating antigen; (3) preparation of monoclonal antibody; and (4) pretreatment and detection of a sample. According to the time and fluorescence resolution fluorescence immunoassay kit for detecting the aflatoxins M1, the detecting time is short, the average recovery rate is high, the pretreatment of the sample is simple, site operation detection can be achieved, the application is wide, and the detection cost is low; and the time and fluorescence resolution fluorescence immunoassay kit for detecting the aflatoxins M1 synchronously has the advantages that the detecting specificity is high, the intra-batch and inter-batch differences are small, the sensitivity is high and the operation is simple and rapid, the time and fluorescence resolution fluorescence immunoassay kit for detecting the aflatoxins M1 is particularly suitable for detection of large scale of samples and the like.

The invention relates to a Bacillus pumilus for degrading aflatoxin M1, collected under CGMCC NO. 10228 and active proteins secreted by the Bacillus pumilus. The Bacillus pumilus is obtained by separating and screening from excrement of herbivores, and the active proteins are extracellular active proteins. The invention also provides a bacterial culture solution of the Bacillus pumilus or a method of using the active proteins secreted by the Bacillus pumilus to degrade aflatoxin M1, and application of the method. By using the method, the bacterial culture solution of the Bacillus pumilus has a degrading rate for aflatoxin M1 not less than 90% in 12 h at 37 DEG C, and the active proteins have a degrading rate for aflatoxin M1 not less than 90% under the same conditions. The Bacillus pumilus and the active proteins secreted thereby have high detoxification activity, high specificity, high acting speed and mild conditions and are applicable to degrading and eliminating aflatoxin M1 in animal-origin foods, and the Bacillus pumilus has the advantages of high bioactivity and probiotic performance.

The invention discloses a method for removing aflatoxin M1 in a liquid system by using lactobacillus rhamnosus, and relates to a detoxifying method for toxins. The method comprises the following stepsof (1) pretreating lactobacillus rhamnosus; (2) regulating the pH of the liquid system containing the aflatoxin M1; and (3) adding the pretreated lactobacillus rhamnosus obtained in the step (1) to the liquid system obtained in the step (2), performing shaping cultivation, and separating the lactobacillus rhamnosus from the liquid system, so that the liquid in the aflatoxin M1 is removed. Throughthe method provided by the invention, the aflatoxins M1 in the liquid system can be efficiently removed, and the technical problems existing in the prior art are solved. The method is environment-friendly and simple to operate.

The invention discloses a time fluorescence resolved fluorescence immunoassay kit for detecting aflatoxin M1 in milk. The time-resolved fluorescence immunoassay kit for aflatoxin M1 in milk is composed of a porous coated plate, buffer solution, aflatoxin M1 standard substance, antibody freeze-dried substance of the aflatoxin M1, europium labelled Anti-MIgG, washing liquid and enhancement solution.A detection method of the time fluorescence immunoassay kit for detecting the aflatoxin M1 comprises the following steps: (1) preparation of immunogen; (2) preparation of coating antigen; (3) preparation of monoclonal antibody; (4) pretreatment and detection of a sample. The kit provided by the invention needs short detection time, has high average recovery rate, is simple in pretreatment of thesample, can be operated on site for detection, can be applied widely, needs low detection cost, and simultaneously has the advantages of strong detection specificity, small within-run and between-rundifference, high sensitivity, being simple and fast to operate, and particularly being suitable for detection of the samples in large scale.