1232 results about "Elisa method" patented technology

The vascular endothelial growth factor (VEGF) activity in a patient's bloodstream or other biological sample can serve as a diagnostic and prognostic index for cancer, diabetes, heart conditions, and other pathologies. Antibody-sandwich ELISA method and kits for VEGF as an antigen were developed to detect VEGF levels in biological samples from animal models and human patients and are used as a diagnostic / prognostic index.

The presence and quantity of an antibody of interest in a patient's bloodstream or other biological sample can serve as an important clinical or other analytical or diagnostic tool. ELISA methods, and kits for such assays, as well as anti-idiotypic antibodies and hybridomas producing them, are developed to detect levels of the antibody in biological samples, which are from, for example, animal models and human patients.

The invention discloses an O type foot-and-mouth disease 146S antigen quantitative ELISA (enzyme-linked immuno sorbent assay) detection kit and a method for using the same. The kit comprises an ELISA plate, an O type foot-and-mouth disease standard reference antigen, a demulsifier, an O type foot-and-mouth disease rabbit antiserum, an O type foot-and-mouth disease guinea pig antiserum, a rabbit anti-guinea pig-horse radish peroxidase conjugate, a guinea pig antiserum dilute solution, a 25-fold PBST (phosphate buffer solution tween) concentrated solution, a carbonate buffer solution capsule, a citric acid-phosphate buffer solution tablet, an OPD (o-phenylenediamine) tablet, a stop solution, a plate sealing membrane, a moving liquid tank and a 96-mesh U-shaped dilution plate. The kit is an organic combination of a sucrose density gradient centrifugation method and an indirect sandwich ELISA method, integrates the advantages of the sucrose density gradient centrifugation method and the indirect sandwich ELISA method, is simple to operate and good in stability, is suitable for batch detection, can be used for distinguishing serum types, and is an ideal substitution method for antigen quantitative and vaccine efficacy detection.

The present invention discloses a chloramphenicol chemiluminescence enzyme-linked immunodetection kit, which comprises a kit body, an enzyme label plate placed inside the kit body, and reagents placed inside the kit body, and is characterized in that every hole of the enzyme label plate is coated with coating antigen, the coating antigen is a chloramphenicol and carrier protein conjugate, and the reagents comprise horseradish peroxidase-labeled chloramphenicol monoclonal antibody, a series of chloramphenicol standard solutions, a concentrated phosphate buffer, a concentrated washing solution and a chemiluminescence solution. The chloramphenicol chemiluminescence enzyme-linked immunodetection kit has characteristics of high sensitivity, simple and rapid detection, and high accuracy, provides a substantially reduced operation time compared to the conventional colorimetric ELISA method, and can be used for detection of chloramphenicol residues in animal tissues (pork, chicken, pork liver and chicken liver), aquatic products (fish and shrimp) and milk.

The invention discloses a kit for detecting pig pseudorabies virus antibodies and a block enzyme-linked immuno sorbent assay (ELISA) method. The kit for detecting pig pseudorabies virus antibodies comprises pig pseudorabies virus monoclonal antibodies which are labelled by horseradish peroxidase, wherein the pig pseudorabies virus monoclonal antibodies are monoclonal antibodies obtained by pig pseudorabies viruses as immunogens and the pig pseudorabies viruses are pseudorabies virus strain Ea. The kit for detecting pig pseudorabies virus antibodies also comprises an enzyme label plate, a sample diluent, negative and positive contrasts, a coloured solution, a washing solution, and a stopping solution. The block ELISA method comprises the following steps of 1, taking out a detection plate pre-coated with virus antigens from the kit for detecting pig pseudorabies virus antibodies, adding diluted blood serum needing to be detected into the detection plate pre-coated with the virus antigens, and simultaneously, setting negative and positive contrast apertures, 2, shaking up the diluted blood serum in the negative and the positive contrast apertures, shaking off a solution in the negative and the positive contrast apertures, and washing the detection plate by the washing solution, and 3, adding the pig pseudorabies virus monoclonal antibodies labelled by horseradish peroxidase into the negative and the positive contrast apertures, washing, adding the colored solution into the negative and the positive contrast apertures to carry out room-temperature coloration in the dark, adding the stopping solution into the negative and the positive contrast apertures, and determining OD630nm values of the negative and the positive contrast apertures by an ELISA apparatus. The block ELISA method has the advantages of good singularity, high sensitivity, short detection time, and high accuracy because of utilization of an S / N ratio method in result determination.

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 a protein chip kit for detecting inflammatory factors, which comprises a basement membrane (1) and a reactant and a detection agent (2). A plurality of kinds of specific antibodies are fixed on the basement membrane; the specific antibodies and the inflammatory factors can undergo an antigen-antibody reaction and each specific antibody is respectively fixed on the basementmembrane for forming a plurality of independent recognition loci; and the reactant and the detection agent are used for detecting whether substances capable of undergoing the antigen-antibody reaction with the specific antibodies exist in a sample to be detected or not by an array-ELISA method. The invention also discloses a method for preparing the protein chip kit for detecting the inflammatoryfactors, which comprises a step of fixing the specific antibodies on the basement membrane. The kit of the invention adopts protein chip technology, can detect 40 inflammatory factors, overcomes a plurality of shortcomings and has the advantages of low cost, convenient use, high sensitivity and accuracy, high flux, small using amount of samples, capability of being popularized in an ordinary laboratory, massive production and the like.

The invention discloses a reagent kit and an enzyme-linked immunochromatography for detecting various organophosphorus pesticide residues and particularly relates to the reagent kit and the enzyme-linked immunochromatography for detecting organophosphorus pesticide residues in the production of vegetative agricultural products. The enzyme-linked immunochromatography for detecting the organophosphorus pesticide residues uses the high-sensitivity characteristic of enzyme to carry out detection by adopting the immunochromatography; compared with a traditional enzyme-linked immunoserbent assay (ELISA) for detecting organophosphorus pesticides, the enzyme-linked immunochromatography has the advantages of simplicity and convenience in operation, and visual result, and no special instrument equipment is needed; and compared with a colloidal gold immunochromatography, the enzyme-linked immunochromatography has the advantages of high sensitivity, strong specificity, accuracy in quantification by combining with a readout instrument and the like.

The invention discloses double-antibody sandwich ELISA (enzyme linked immunosorbent assay) kit used for detecting an avian leukosis group specific antigen. The kit comprises an enzyme plate coated by a monoclonal antibody which is secreted by a hybridoma cell strain the preservation serial number of which is CGMCC (china general microbiological culture collection center) No.5961. The invention also discloses a double-antibody sandwich ELISA method which is established by utilizing the monoclonal antibody and is capable of rapidly and effectively detecting an ALV (avian leukosis virus). In the double-antibody sandwich ELISA method, the monoclonal antibody prepared by pronucleus expressive HLJ09mdj-1p27 albumen is utilized as a peridium antibody, and antibodies prepared by p27 are utilized as a detection antibody. According to the invention, the minimum detection amount of the p27 is 1.25 ng / ml, the method is not reacted with the common virus of birds, and the specificity is good. The method is utilized to detect egg white and an anus swab sample, and the coincidence rate is respectively 96.5% and 88.9% compared with a PCR (polymerase chain reaction) method; and the result proves that the method has the advantages of convenience, celerity, differentia, sensitivity and the like, and is useful for the detection and population purification of the ALV.

This invention relates to a rapid and efficient method for carrying out enzyme-linked immunosorbent assay for detection of minute quantities of biomolecules such as antigen, antibody etc. This invention particularly relates to microwave mediated immobilization of antigen or antibody on to the activated surface followed by performing subsequent steps of ELISA by controlled microwave irradiation. The invented procedure has dramatically reduced the total time required for ELISA to less than 10 minutes from hours to days. The invented ELISA procedure is rapid, economical, reproducible and simple and can be automated. The invented procedure is useful for carrying out ELISA in clinical diagnostics, molecular biology, agriculture, sericulture, food technology, environmental science, biomedical research and other related fields.

The invention relates to an anti-Golgi protein antibody and an application thereof. The invention recombines human GP73 protein immunity animal to obtain an anti-GP73 polyclonal antibody and a monoclonal antibody which specifically aims at GP73 and builds a plurality of methods for detecting GP73 in clinical tissue sections and serum samples, such as immunohistochemical stain and double antibody sandwiched ELISA method and the like. Tests prove that the polyclonal and monoclonal antibodies can be used for preparing a plurality of GP73 detecting agents of different detecting methods.

The invention relates to a group of anti-human CD146 molecules which are developed by utilizing the biological technology and an established high-sensitive sandwich ELISA method for detecting the solubility CD146. The invention includes anti-human CD146 mouse monoclonal antibodies of AA1, AA2, AA3, AA4, AA5 and AA7, and the method of utilizing the combination of the antibodies and the sandwich ELISA specificity for detecting the solubility CD146. The group of antibodies can identify the human source CD146 on the molecular, the cellular and the tissue levels, which can be divided into two categories based on the identified different epitopes. The sandwich ELISA method which is combined by the antibody AA1 and another strain of anti-human CD146 mouse monoclonal antibody AA98 can detect the solubility CD146 at each milliliter nano-gram level. The antibodies and the detection means can become the effective detection or diagnosis tools and method in the basic research or the clinical application and provide good carriers for the targeted therapy of the CD146-related diseases at the same time.

The invention relates to a competitive ELISA method based on a foot-and-mouth disease A type VP1 protein and its monoclonal antibody, also relates to a preparation method of the foot-and-mouth disease A type VP1 protein, and a preparation method of the monoclonal antibody of the foot-and-mouth disease A type VP1 protein, and belongs to the technical field of animal immunological detection. In the invention, a primer pair C1 and C2 and a primer pair E1 and E2 are amplified to obtain a gene sequence of the foot-and-mouth disease A type VP1 protein, the foot-and-mouth disease A type VP1 protein is obtained by constructing an expression plasmid, introducing the expression plasmid into a prokaryotic expression host and carrying out inducible purification, the foot-and-mouth disease A type VP1 protein monoclonal antibody is obtained by treating the foot-and-mouth disease A type VP1 protein as an antigen through a hybridomas technology, and the competitive ELISA method used for detecting a foot-and-mouth disease A type antibody is established based on the foot-and-mouth disease A type VP1 protein and its monoclonal antibody. The detection method has a strong specificity and a good stability, and can be used for detecting a foot-and-mouth disease A type serum antibody. By comparing a result obtained through the detection method with a liquid phase blocking ELISA kit, the coincidence rate is 95.8%.

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

The invention discloses a process for preparing variable zone genes of high affinity tumor necrosis factor (TNF) resistant monoclonal antibody (F6 mAb) comprising, using recombinated human TNF immune BALB / c mouse to prepare a plurality of mouse Anti-TNF monoclonal antibody, screening high affinity F6 mAb using indirect ELISA method. By cloning the monoclonal antibody light chain and heavy chain variable zone genes, the monoclonal antibody light chain and heavy chain variable zone gene sequence and amino acid sequence can be obtained, and the unicity of the gene sequence and protein sequence can be confirmed.

This invention discloses one enzyme immune test method and agent case for 3- methyl quinoline 2-carboxyl acid in immune chemical analysis technique field, which comprises the following steps: processing immune antigen and cover antigen and alloantibody and pre-processing sample and establishing ELISA method; the matched agent case is composed of 3- methyl quinoline 2-carboxyl acid alloantibody covered by MQCA and egg albumin couple and MQCA standard sample; the sample is processed through metaphosphoric acid to release MQCA and MAX and indirectly competition ELISA method.

The vascular endothelial growth factor (VEGF) activity in a patient's bloodstream or other biological sample can serve as a diagnostic and prognostic index for cancer, diabetes, heart conditions, and other pathologies. Antibody-sandwich ELISA methods and kits for VEGF as an antigen are provided to detect types of VEGF levels in biological samples from animal models and human patients and can be used as a diagnostic / prognostic index.

The invention discloses a hybridoma cell strain capable of secreting monoclonal antibodies to quinolones and application of the monoclonal antibodies thereof. Ciprofloxacin (CIP) coupled with bovine serum albumin is used as an antigen to immunize BALB / c mice and cell fusion, screening and cloning are carried out so as to obtain one hybridoma cell strain 1F1 capable of stable passage and secretion of monoclonal antibodies (MAb) to quinolones, wherein, the accession number of the hybridoma cell strain 1F1 is CGMCC No. 5608. The titres of ascitic fluids of the 1F1 monoclonal antibodies are up to 10<-7>, and the type and the subclass of the monoclonal antibodies are IgG1 and kappa chain. According to indirect competitive ELISA analysis, the 1F1 monoclonal antibodies perform specific reactions to quinolones like ciprofloxacin, enrofloxacin, ofloxacin, danofloxacin, norfloxacin, enoxacin, marbofloxacin, sarafloxacin and difloxacin. An ELISA method, a kit and test paper for detecting residual of quinolones in food are developed by using the 1F1 monoclonal antibodies.

The present invention provides a disc-like assay chip, and more specifically, the present invention provides an assay chip which can effectively clean a cleaned object existing in a fluid loop, and can be appropriately applied to a detection method of a reaction system using a step of necessarily cleaning the cleaned object, such as an immunofluorescent antibody method, and applied to a detection method using an enzymatic reaction (especially, an ELISA method). The present invention further provides a disc-like assay chip which is mounted on a centrifugal device such as a rotational disc and can perform detection or quantitation on a target substance through optical measurement after a specimen and a reagent react by use of a centrifugal force generated by rotation of the centrifugal device.

The invention discloses a magnetic particle chemiluminescence quantitative assay kit for anti-ribosomal P protein antibody IgG. The kit includes: anti-ribosomal P protein antibody IgG calibrator; Ribosomal P protein antigen and bovine serum albumin-labeled Tris buffer; alkaline phosphatase-labeled goat anti-human polyclonal antibody and bovine serum albumin in Tris buffer; streptavidin-labeled magnetic particles and bovine serum albumin Serum albumin in Tris buffer; wash solution. Based on the traditional membrane strip immunoassay and enzyme-linked immunosorbent assay, the detection method of the kit increases the sensitivity and linear range by 3-5 orders of magnitude, and realizes quantitative detection in a real sense, with rapid response, reliable results, and It can be used in conjunction with a fully automatic chemiluminescence immunoassay analyzer to realize fully automatic use, and has irreplaceable important value for clinical diagnosis.

The invention discloses a double-antibody sandwich method for detecting salmonella typhimurium in food based on monoclonal antibodies, belonging to the technical field of immunoassay. Salmonella typhimurium ATCC13311 and smooth salmonella typhimurium LPS are adopted for mixed immunity of a 7-week BALB / c mouse, 10 LPS monoclonal antibodies are obtained by immunity, fusion and screening, horse radish peroxidases (HRP) are labeled respectively, and the salmonella typhimurium is paired two by two. A sandwich enzyme-linked immuno sorbent assay (ELISA) method is established by taking 6E2 CGMCC No.7206 monoclonal antibodies as coated antibodies and enzyme-labeled antibodies and by taking the salmonella typhimurium as standards, and the LOD is 500cfu / mL. The sandwich method, established by using the monoclonal antibodies which are highly uniform in physicochemical property and high in specificity and can be prepared on a large scale, is high in sensitivity and low in cost; the salmonella typhimurium is not in cross reaction with salmonella enteritidis, salmonella arizonae, E.coli, E.coliO157:H7, enterobacter sakazakii, staphylococcus aureus and listeria monocytogenes; a quick and efficient analysis way is provided for detection of the salmonella typhimurium in the food.

The 19 monoclonal antibody strains of 8 monoclonal antibody for 8 plant viruses including broad bean wilt virus, cucumber mosaic virus, tomato mosaic virus, cane mosaic virus, etc are disclosed. The high-specificity high-sensitivity ACP-ELISA and TAS-ELISA methods for detecting relative viruses are disclosed on the basis of said 8 antibodies.

The invention relates to a method for external detection of polymerizing power of disease-related protein promoted by body fluids of subjects. More specifically, the invention concerns a method for detecting the oligomer formation power of disease-related protein such as alpha-synuclein promoted by body fluids of the subjects in vitro. The method comprises: incubating purified recombinant human disease-related protein and body fluids of the subjects by stirring at a certain temperature to make the two components be polymerized, and detecting the content of oligomers of disease-related protein such as alpha-synuclein in the incubated body fluids by utilizing a method that can detect protein oligomers. The method provided by the invention has advantages that: 1) the need for body fluid sample is less, so the subjects are easy to accept; 2) operations are simple; and 3) the result obtained from the detection method reflects the synthetic action of various factors of polymerization of disease-related protein to subjects, and the result is particularly more favorable for determining comprehensive pathophysiological situations of patients with Parkinson disease caused by complex factors.

The present invention discloses a sulfanilamide drug chemiluminescence enzyme-linked immunodetection kit, which comprises a kit body, an enzyme label plate placed inside the kit body, and reagents placed inside the kit body, and is characterized in that every hole of the enzyme label plate is coated with coating antigen, the coating antigen is a sulfanilamide mother nucleus and carrier protein conjugate, and the reagents comprise sulfanilamide monoclonal antibody, horseradish peroxidase-labeled goat anti-mouse antibody, a series of sulfanilamide standard solutions, a concentrated phosphate buffer, a concentrated washing solution and a chemiluminescence solution. The sulfanilamide drug chemiluminescence enzyme-linked immunodetection kit has characteristics of high sensitivity, simple and rapid detection, high accuracy, and more drug detection types, provides a substantially reduced operation time compared to the conventional colorimetric ELISA method, and can be used for detection of residues of the 17 sulfanilamide drugs in animal tissues (pork, chicken, pork liver and chicken liver), aquatic products (fish and shrimp), eggs, milk and milk powder.

The invention provides a clenbuterol (CLB) magnetic particle separation enzyme-linked immunosorbent assay (ELISA) method, belonging to the technical field of food safety immunodetection. The method comprises the following steps: the immunodetection principle of the competition method is adopted to connect CLB with bio-enzyme and prepare an enzyme labeled antigen reagent, antibody against fluorescein isothiocyanate (FITC) is absorbed on the surface of magnetic particles to prepare a magnetic separation reagent, FITC is connected with the antibody against CLB to prepare an antibody reagent; CLB in a sample and enzyme-labeled CLB compete to combine with a small quantity of FITC-labeled antibody against CLB and form an antigen-antibody composite; after the magnetic separation reagent is added, the antibody against FITC connected with the surface of magnetic particles captures the composite on the surface of magnetic particles; and washing, and finally adding substrate to detect. The invention has the following advantages: (1) magnetic particles replace the traditional ELISA plate to be used as solid carrier and ensure that the immunoreaction is performed more fully and fast under the near-liquidus condition; compared with the traditional ELISA method, the method is characterized by high specificity and good repeatability; and (2) the one-step competition method principle is adopted, thus the detection time is short.

The invention provides a multi-antigen enzyme linked immunosorbent assay (ELISA) kit for detecting an African swine fever virus (ASFV) antibody, belonging to the field of a biotechnology and diagnosis and research of animal-borne diseases. The multi-antigen enzyme linked immunosorbent assay kit comprises expression and purification of three types of ASFV recombined antigens, preparation of positive and negative control blood serum of an ASFV antibody, optimal envelope antigen combination and concentration determination, optimization of multi-antigen ELISA (MA-ELISA (Microalbumin-Enzyme Linked Immunosorbent Assay)) reaction parameters, determination of an ASFV antibody negative blood serum critical value, MA-ELISA detection artificial infection and determination of sensitivity, specificity and repeatability of field blood serum samples. By detecting and testifying a large quantity of known blood serum samples, the sensitivity, the specificity and the repeatability of detecting the ASFV antibody by the MA-ELISA are obviously higher than those of an ELISA method recommended by World Organization for Animal Health and oversea similar kits; and the multi-antigen enzyme linked immunosorbent assay kit can be used for ASFV serological diagnosis, epidemiological investigation and live pig import and export quarantine inspection.

The invention discloses a preparation method and application of a product obtained by coupling melamine with carrier protein, as well as a melamine antibody. The product obtained by coupling the carrier protein with the melamine is used as artificial antigen and applied to an immunological method for melamine detection. The preparation method comprises the following steps of immunizing animals with the coupled product so as to prepare an antibody used for melamine detection, fusing BALB / C mouse spleen cells immunized with the coupled product and SP2 / 0 mouse myeloma cells, obtaining hybridoma capable of stably transferring culture and secreting anti-melamine specific monoclonal antibodies by screening positive hybridoma and cloning cells, and preparing an ascites monoclonal antibody. The prepared monoclonal antibody is utilized to establish a direct competitive ELISA method having high specificity, sensitivity and accuracy to the melamine, as well as an immune colloidal gold test strip. The preparation method for the product obtained by coupling melamine with carrier protein, as well as the melamine antibody provides service for the rapid detection of melamine-type residue in foods.

The invention belongs to the field of bioengineering and provides an Echinococcus granulosusglutathione transferase gene a base sequence of which is shown in SEQ ID NO: 1. The invention also provides a protein encoded by the Echinococcus granulosusglutathione transferase gene. The amino acid sequence of the protein encoded by the Echinococcus granulosusglutathione transferase gene is shown in SEQ ID NO: 2, or is the same as 1st-219th amino acid sequences as shown in SEQ ID NO: 3. The recombinant protein can be used for immunodiagnosis of cystic echinococcosis patients and can be used for coating a plate; and the serums of the patients having cystic echinococcosis, alveolitoid echinococcosis, cysticercosis and other parasite diseases and healthy people are detected by using an enzyme-linked immunosorbent assay (ELISA), and the obtained sensitivity and specificity are respectively 72.41% and 92.36%.