552 results about "Enzyme linked immunoassay" patented technology

The invention provides an aflatoxin B1 (AFB1) magnetic separation enzyme-linked immunity quantitative detection method, belonging to the field of food safety immunoassay technique. The method adopts the immuno-detection principle of competition law; and AFB1 is connected with biological enzyme to prepare enzyme-labeled antigen reagent, anti-fluorescein isothiocyanate (FITC) antibody is absorbed onthe surfaces of magnetic particles to prepare magnetic separation reagent, and the FITC is connected with the AFB1 antibody to prepare anti-reagent. In a sample, the AFB1 competes with the enzyme-labeled AFB1 and is combined with a small amount of FITC-labeled anti-AFB1 antibody, so that antigen-antibody complex can be formed. After the magnetic separation reagent is added, the complex is caughtonto the surfaces of the magnetic particles by the anti-FITC antibody connected on the surfaces of the magnetic particles. After being washed, the product is finally added with substrate and detected.The method has the advantages that (1) the magnetic particles are used for replacing the traditional enzyme-labeled plate to be taken as a solid-phase carrier, so that immunoreaction is carried out under the approximate liquid phase condition; and the reaction is more complete and rapid, and has the characteristics of high specificity and good repeatability compared with the traditional enzyme-linked immuno sorbent assay (ELISA); furthermore, (2) by adopting one-step competition law principle, the time used for detection is short.

The invention discloses an enzyme immunoassay of testing the bice green residues in animal derived food, which comprises an enzyme label plate covering bice green antigen, enzyme label bice green antibody working solution, bice green standard solution, substrate solution, substrate buffer solution, reaction termination solution, concentration washing liquid and sample dilute solution. The invention further discloses a method for using the immunoassay to test bice green residues, which comprises sample pretreatment, testing via the immunoassay, processing and analyzing result. The inventive immunoassay of bice green test uses direct competition enzyme-linked immunoassay adsorption analysis technique, with high sensitivity, high stability, simplified operation, reduced reaction time, reduced error caused by complex operation, reduced cost, wide application for testing samples and high practicality.

The invention discloses a method for detecting a sulfanilamide medicine and a special enzyme-linked immunoassay reagent kit thereof. The enzyme-linked immunoassay reagent kit comprises a sulfanilamide medicine monoclonal antibody and the sulfanilamide medicine. The sulfanilamide medicine monoclonal antibody is secreted by the hybridoma cell line SAs of a sulfanilamide monoclonal antibody, whose preservation number is CGMCC No. 3393. In the enzyme-linked immunoassay reagent kit of the invention, an indirect competition ELISA (Enzyme-Linked Immuno Sorbent Assembly) method is mainly used for qualitatively or quantitatively detecting the content of the sulfanilamide medicine in products (especially milk, pork, chicken, eggs, honey, fish, shrimp and the like) eaten by animals or people. The reagent kit and the detection method of the invention have low requirements on the pretreatment of samples and simple pretreatment process of the samples and can detect mass samples quickly at the same time. By using the sulfanilamide medicine monoclonal antibody with high specificity, the detection method is convenient and simple, and the invention has the characteristics of high specificity, high sensibility, high precision, high accuracy and the like.

The invention discloses a method for detecting the specific antibody of classical swine fever virus and a special ELISA kit thereof. The kit includes a classical swine fever virus antigen and an enzyme-labeled classical swine fever virus single-epitope specific antibody; the said swine fever virus antigen is a polypeptide containing one or more than one amino acid residue sequence described in sequence 1. The detection reagent of the classical swine fever virus specific antibody of the present invention can carry out effective detection to the classical swine fever virus specific antibody by solid-phase antigen competition ELISA (blocking method); The B cell epitope ensures the differential diagnosis, and the high degree of conservation of the epitope among various strains ensures the specificity of detection.

The invention discloses a kit for rapid detection of staphylococcus aureus in a sample and a detection method thereof, belonging to the technical field of immunological detection. In the invention, a staphylococcus aureus immunogen inactivated with formaldehyde is used for immunizing a healthy New Zealand rabbit to obtain a polyclonal antibody to serve as a coated antibody, and used for immunizing a BALB/C mouse and performing cell fusion to obtain a monoclonal antibody to serve as a secondary antibody, thus, the kit for performing a double antibody sandwich enzyme-linked immunosorbent assay on the staphylococcus aureus in food (milk) is established, and a rapid and efficient detection means is provided for residual detection of the staphylococcus aureus in the food, and the advantages of lower cost and better stability and repeatability are achieved. A detection limit of the kit is 105cfu/mL and is suitable for detecting mass samples.

A conjugate of norfloxacin is prepared from norfloxacin semi-antigen and the bovine serum albumin (or egg albumin) as the carrier able to generate immunogenicity through coupling. It can be used for preparing the reagent kit for the enzyme-linked immunoassay of norfloxacin.

The invention provides an enzyme-linked immunoassay method for determining the content of a cell DNA (Deoxyribonucleic Acid) injury marker H2AX. The enzyme-linked immunoassay method is characterized by comprising the steps that sandwich recognition on the specificity of a target protein H2AX is carried out by using a specific H2AX antibody and an enzyme-linked second antibody, a substrate after enzyme reaction is added, then the substrate is subjected to enzyme catalysis to be changed into a fluorescence product, wherein quantitative determination for the H2AX content can be realized according to a fluorescence value of the substrate. According to the method, the content change of the H2AX in the cells, caused by the fact that cigarette smoke is exposed, is quantitatively determined so that the aim of evaluating the virulence of cigarette smoke genes is realized. Compared with a traditional organic dye, the enzyme-linked antibody has the advantages that the catalyzing frequency of an enzyme is high, an amplification reaction effect is amplified and the detection sensitivity is improved; the enzyme has a stable property at a room temperature, is low in price, and is suitable for a lot of analysis. An enzyme-linked immunosorbent assay has the advantages of rapidness, accuracy, high throughput, high sensitivity and the like.

The invention discloses an indirect ELISA kit for detecting an African swine fever virus antibody and an application thereof, and belongs to the technical field of biology. The kit adopts prokaryotic expression recombinant P30 protein as a coating antigen, and detects the antibody of African swine fever virus in porcine serum based on the indirect ELISA principle. The coating antigen in a 96-well plate of the kit is prokaryotic expression recombinant P30 protein which has good antigenicity. The enzyme-linked immunoassay kit provided by the invention comprises a 96-well plate coated with P30 protein, a positive control, a negative control, a horseradish peroxidase-labeled rabbit anti-porcine IgG polyclonal antibody, a concentrated washing liquid, a serum diluent, a TMB substrate, and a terminating liquid. The kit of the invention is applicable to the screening of large quantities of samples, and main reagents in the kit are provided in a form of operating fluid which is convenient for use.

The invention provides an electrochemical DNA biosensor for detecting PML/RAR alpha fusion gene of acute promyelocytic leukemia. The gene to be detected is the PML/RAR alpha fusion gene of the acute promyelocytic leukemia (APL); and the method comprises the following steps: (1) designing and synthesizing a specific sequence of the APL according to the selected fusion sites of a universal primer sequence of the gene type of the APL to be detected; and (2) constructing a nano modified electrode by a Au nano material, and constructing the nano electrochemical biosensor used for detecting the PML/RAR alpha fusion gene of the APL by the electrochemical enzyme-linked immunoassay technology. The method greatly improves the sensitivity of the biosensor so as to realize early diagnosis of the acute promyelocytic leukemia.

The invention belongs to the technical field of immunoassay and diagnosis and relates to a method for magnetic antibody immunoassay chemiluminescence detection of treponema pallidum antibodies. The method comprises the following steps of: using GoldMag particles as carriers, coating one or more antigens of specific recombinant proteins of treponema pallidum antibodies, TP15, TP17, TP47 and TP44.5, then adding a sample to be detected and enzyme labeled antigens, and finally adding luminescence substrates for luminescence detection. The GoldMag particle surface has a larger coupling capacity to the antigen and has characteristics of high specificity and no pollution of the enzyme linked immunoassay as well as high sensitivity of chemiluminescence, and therefore, the method has the advantages of high detection sensitivity, good specificity, wide linear range, no radioactive pollution and the like.

The invention discloses an enzyme-linked immunoassay kit for acid orange II, which belongs to the technical field of enzyme-linked immunosorbent analysis. Reagents in the kit provided by the invention comprise an enzyme label plate coated by a coupled substance of the acid orange II serving as a coating antigen and a carrier protein, standard solution of the acid orange II, solution of enzyme labeled goat-anti-rabbit antigen, antibody solution of the acid orange II, luminous solution, washing solution, coating solution and closed solution. The carrier protein of the coupled substance is bovine serum albumin or egg albumin with a molecular weight of 6.7 to 6.8 KDa. The coating antigen of the coated enzyme label plate is prepared by coupling the acid orange II and egg albumin, and an artificial immunogen for preparing the angiten is prepared by coupling the acid orange II and the bovine serum albumin. The maximum acid orange II detection range of the kit is 0.1 to 10 ng/mL. The enzyme-linked immunoassay kit of the invention has the characteristics of simplicity, quickness, accuracy and high sensitivity and can play an important role in the acid orange II detection of synthetic non-edible pigment in foods (such as marinated products and cayenne pepper) and drinks.

The invention provides a kit for detecting a novel coronavirus neutralizing antibody by enzyme-linked immunosorbent assay and a detection method thereof, the kit comprises two ELISA plates coated withantigens, one ELISA plate is coated with an RBD antigen of S protein, the other is coated with NTD of S protein; and the two ELISA plates are respectively sealed by using sealing liquid. The kit disclosed by the invention is used for detecting the novel coronavirus neutralizing antibody and evaluating the immune efficacy of the vaccine. The kit for detecting the novel coronavirus neutralizing antibody by the enzyme-linked immunosorbent assay provided by the invention applies the enzyme-linked immunosorbent assay with high sensitivity, good specificity and good repeatability, is beneficial towashing and removing non-specific binding and separating unreacted substances by immobilizing the antigen, has the characteristics of high sensitivity and good specificity, and is suitable for accurate detection of high-throughput samples.

The invention relates to the field of medical biotechnology, in particular to a making method for coronary heart disease and apoplexy risk diagnosis kit. The kit is used for quantitative analysis of Lp-PLA2 level in human plasma or blood serum by enzyme linked immunosorbent assay, so as to prevent heart disease and apoplexy.

The invention relates to a method for preparing a porcine circovirus type 2 colloidal gold antibody fast test strip. In the method, a porcine circovirus type 2 ORF2 protein is expressed by utilizing genetic engineering, and the test strip is prepared by the principle of enzyme-linked immunoassay and membrane chromatography to fast test antibodies in the blood or serum of pigs. The test strip can be widely used for clinically testing porcine circovirus diseases, has no cross reaction with other viruses, and obtains test results 98.63 percent and 95.83 percent consistent with those obtained by the two methods of the ELISA and neutralization test. Compared with the ELISA, a recombinant antigen immune colloidal gold has the remarkable advantages of high security, no need of culturing the viruses per se, the avoidance of virus spread caused by the operation of the viruses, large-batch preparation, simple process, low production cost, stable and homogeneous antigen components, simple, convenient and labor-saving operation, no apparatus, high detection result specificity, high repeatability, the short time of 15 minutes for the whole test, simple, convenient, fast and accurate operation, high sensitivity, intuition and easy result judgment.

The invention belongs to the veterinary medicine residual analysis and immunity analysis technique field and specifically relates to an enzyme-linked immunity method and the reagent kit thereof which can discern specificity monoclonal antibodies of tylosin and tilmicosin and detect the residuals of tylosin and tilmicosin at the same time; the monoclonal antibodies in the invention is secreted from hybridoma cell strain P3C4 established by the applicant; the hybridoma cell strain is preserved in China Center of Type Culture Collection; the number of preservation of the hybridoma cell strain is CCTCC No: C200719; the invention discloses the preparation method and enzyme-linked immunity detection method of the monoclonal antibody, coating antigen and immunogen. Compared with the prior art, the monoclonal antibody prepared in the invention can discern tylosin and tilmicosin at the same time and add the detecting object in the prior art. The reagent kit and the method in the invention has the advantages of simple, convenience, swiftness, sensitivity and accuracy; furthermore, the reagent kit and method in the invention can detect the residuals of tylosin and tilmicosin in an animal edibility tissue at the same time.

The invention relates to a kit used for rapidly detecting Escherichia coli O157:H7 in a sample, and a detection method thereof. The invention belongs to the technical field of immunological detection. According to the invention, a heated and deactivated Escherichia coli O157:H7 immunogen is used for immunizing a healthy New Zealand rabbit, such that a polyclonal antibody is obtained, and the polyclonal antibody is adopted as a coating antibody; a BALB/C mouse is immunized, and cell fusion is carried out, such that a monoclonal antibody is obtained, and the monoclonal antibody is adopted as a secondary antibody; and a double-antibody sandwich ELISA kit of Escherichia coli O157:H7 in foodstuffs (meat) is established. With the kit, a rapid and highly efficient detection means is provided for the detection of the residue of Escherichia coli O157:H7 in foodstuffs. The kit is advantaged in relatively low cost, relatively good stability, and relatively good repeatability. According to the invention, a detection limit is 105cfu/mL. The kit and the method are suitable for large-batch detections of samples.

Disclosed are a hepatitis C virus antigen enzyme-linked immunoassay reagent box and a method for making the same. The invention obtains the cell strain of excretive anti HCV core antigen by analyzing the core antigen array of the hepatitis C virus different type and cloning the core antigen gene of HCV, purifying out the high activity monoclonal antibody with four core aa expression sites of HCV, wherein the Cab1 and Cabs are used as coating antibodies, the Cab3 and Cab4 are used as enzyme labeled antibodies; employing double antibodies sandwich technology to prepare HCV-cAg ELISA diagnosing reagent box.

The invention relates to a preparation method of a diagnostic reagent kit for early diagnosis and recurrence monitoring, and the like of ovarian cancer. The preparation method comprises the following steps of: the recombination expression of a marker HE4 (Human Epididymis Protein) in eukaryotic cells, monoclonal and polyclonal antibody preparation by HE4 obtained by the recombination expression in the eukaryotic cells, and the diagnostic reagent kit (a double antibody sandwich method and an enzyme-linked immunization method) development by the prepared monoclonal and polyclonal antibodies. The invention can ensure that HE4 obtained by the recombination expression in the eukaryotic cells by utilizing a gene engineering method has the same glycosylation degree with natural HE4; the space conformation thereof more approaches to a natural antigen; the prepared monoclonal and polyclonal antibody is used as a coating antibody, and the other monoclonal antibody is used as an enzyme mark antibody to develop the reagent kit; and the sensitivity and the specificity are higher.

The invention relates to a diagnostic kit for tuberculosis and mycobacterium tuberculosis infectors, a preparation method and an application method thereof. By using the linker for encoding 15 amino acid (G4S1) 3, the encoding genes (SEQ.ID.NO.6) of a mycobacterium tuberculosis specific antigen Rv3875 and Rv3874 are connected in series, and then inserted into an E. coli expression vector, and the high-efficiency expression and purification for the fusion protein of Rv3875 and Rv3874 in the E. coli are achieved. The recombinant protein at least comprises 8 T cell epipositions which can be used for cell immunity diagnosis, and a diagnostic kit and diagnostic method for a whole blood IFN-Gamma release analysis method are established by taking the protein as the basis and combining the human IFN-Gamma enzyme-linked immunoassay technology, and can be used for the early, specific diagnosis and screening of tuberculosis and mycobacterium tuberculosis infectors.

The present invention relates to a tetracycline group antibiotic ELIA kit, belonging to the field of enzymoimmunoanlysis technology. Said invention adopts the enzymoimmunoadsorption, protein coupling and biochemical preparation technique to prepare the invented detection kit. Its key technique lies in that after the tetracycline group is renovated, it is coupled with protein BSA by using succinyl oxide as bridge to prepare coating antigen, the tetracgcline group is coupled with protein BSA by using glutaraldehyde as bridge to synthesize immunoantigen, then the enzymoimmunoadsorption indirect competition method can be used to detect tetracycline group. Its detection range is 1950ng/g-1.90ng/g; its detection time only requires 4 hr.

The invention relates to a clothianidin antigen, an antibody and an application thereof, and belongs to the technical field of immunochemistry analysis. The clothianidin antigen and the antibody of the invention are specially used for clothianidin specific polyclonal antibody preparation, enzyme-linked immunoassay (ELISA), and high-sensitivity and rapid detection of clothianidin residues in environment and agricultural products. A hapten is synthesized by substituting a chlorine atom on a thiazole ring of clothianidin, has a chemical name of 3-(5-((3-methyl-2-nitroguanidine)-yl methyl)thiazole-2-mercapto)propionic acid, and is coupled with bovine serum albumin and ovalbumin to prepare an antigen and an envelope antigen. Newzealand white rabbit is immunized by the immunizing antigen to obtain the specific polyclonal antibody of clothianidin. The established ELISA linear scope is 1.1 microgram/L-2 mg/L, and the detection limit is 1.1 microgram/L. The hapten synthetic technology of the invention is simple and feasible, high in antibody specificity, and suitable for detection and on-site monitoring of mass samples in environment and agricultural products.

The invention provides a method for detecting canine rabies virus antibody (IgG) and a detection kit. The kit is composed of a coated plate and a reagent reaction system, and comprises a rabies virus antigen-coated reaction plate, a standard substance, positive contrast serum, a washing lotion (20X), a sample weak solution, an enzyme-marked combination substance, a developer A, a developer B and a stopping solution. The method is characterized in that the method uses an ELISA method to determine that the content of the canine rabies virus antibody (IgG) reaches a absorbance corresponding to a protective level by detecting the standard substance of the kit, and by compared the absorbance of the sample to be detected with the absorbance of the standard substance, the content of the canine rabies virus antibody (IgG) contained in the sample to be detected is judged whether to reach an immunization protective level. The method and the kit can simultaneously detect a lot of samples, and a detection result is remarkably with a result by a neutralization experiment, has high accuracy degree, is suitable for monitoring an immunization inoculation effect and determining an individual immunization state, and can be used for investigating animal eqpidemic diseases.

The present invention belongs to the field of enzyme-linked immunoassay technology, and is especially specific antibody of provera acetate and its preparation and usage in homogenous or heterogenous enzyme-linked immunoassay. The present invention prepares haptens 3-CMO-MPA, 3-CMO-CMA or 3-CMO-MEGA; couples hapten 3-CMO-MPA with protein to prepare immunological antigen; couples haptens 3-CMO-MPA, 3-CMO-CMA and 3-CMO-MEGA with protein separately to prepare three kinds of envelope antigens. When the immunological antigen is used in immunizing animal, the produced antibody can produce specific reaction with provera acetate and may be used in establishing homogenous or heterogenous enzyme-linked immunoassay method for detecting the content of provera acetate in the sample fast in high sensitivity.

The invention discloses an enzyme-linked immunoassay kit for detecting phenylethanolamine A by a direct competition method. The enzyme-linked immunoassay kit for detecting phenylethanolamine A provided by the invention comprises an enzyme label plate coated by phenylethanolamine A specific antibodies, and a phenylethanolamine A horseradish peroxidase marker. The enzyme-linked immunoassay kit for detecting phenylethanolamine A of the invention is sensitive, rapid, accurate, and is mainly used for screening of large quantities of samples; main reagents in the kit are provided in the form of operating fluid, and the kit is convenient for using, has the characteristics of high specificity, high sensitivity, high precision, high accuracy, and the like, can rapidly detect residual phenylethanolamine A in feed and animal products.