121 results about "Immunosorbent method" patented technology

The invention discloses a latex-enhanced immunoturbidimetric assay kit for neutropil gelatinase-associated lipocalin, and a preparation method thereof. Polyethylene glycol hexamine is selected as theblocking agent of an antibody binding latex particle. The kit used for latex-enhanced immunoturbidimetric assay of the NGAL has the advantages of high detection specificity, high sensitivity and goodstability, and can efficiently detect the NGAL content in urine, plasma and serum, and the detection result is well associated with fluorescence immunochromatography and enzyme linked immunosorbent assay.

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 belongs to the fields of immunology and biotechnology, and particularly relates to a PSMD4 protein ELISA detection kit as well as a detection method and application to serological diagnosis for liver cancer of the kit. According to extensive and deep research of the inventor, the expression level of PSMD4 protein in the serum of liver cancer patients can be detected according to an enzyme-linked immunosorbent assay (ELISA) method, and the PSMD4 protein is proved for the first time in the serum of the liver cancer patients. The kit comprises an ELISA plate coated with a PSMD4 antibody, a PSMD4 antigen detection antibody, an ELIAS secondary antibody, standard protein and the like. The invention further provides the detection method and application of the kit. The kit is simple and convenient to operate, can detect the content of the PSMD4 protein in the serum of a liver cancer patient correctly and highly sensitively, and provides a novel method for clinical examination and basic research.

The invention discloses a method for detecting mite allergen specific antibody in blood serum. The method comprises the following steps of: coupling mite allergen with surface carboxyl modified magnetic microspheres to prepare immunomagnetic microspheres; and mixing and incubating the blood serum to be detected and the immunomagnetic microspheres in holes of an elisa (enzyme-linked immunosorbent assay) plate to combine mite allergen with mite allergen specificity IgE in the blood serum, and detecting by adopting an enzyme-linked immunosorbent method after the elisa plate is washed. In the method disclosed by the invention, a mite allergen is directly coupled onto the surfaces of magnetic microspheres, thus all the impurities except the mite allergen specific antibody in the blood serum can be conveniently removed; and targeted mite allergen specificity antibody detection is carried out, detection time is shortened, and sensitivity, specificity and accuracy of detection are improved. The mite allergen immunomagnetic microspheres prepared by the invention can be used in standard automatic detection to improve detection efficiency.

The invention relates to a stem tip detoxification and cultivation method for a sweet potato virus seedling. The method comprises the following steps: (1) taking a sweet potato block infected by sweet potato feathery mottle virus and/or sweet potato G virus; cultivating the sweet potato block indoors to sprout; (2) shearing scissoring terminal buds growing to 1.5-2.0 cm for disinfection; (3) peeling off the stem tips with two leaf primordiums in a sterile manner among the terminal buds as explants for vaccination cultivation to obtain regeneration plants; (4) conducting 2-3 times of successive transfer cultivation on the regeneration plants obtained in the step (3) to build a strain; conducting inspection by adopting an indicator plant method and a cellulose nitrate membrane ELISA in sequence to obtain a detoxified strain; (5) proliferating the detoxified strain quickly to obtain the detoxification seedling. The method can quickly build cultivation system suitable for stem tip cultivation and rapid propagation detoxification seedlings of majority of sweet potato varieties/strains.

The invention provides a method for removing PVS viruses of potato test-tube plantlets. The method comprises the steps of stripping the stem tips of the pretreated test-tube plantlets and then putting the test-tube plantlets into a stem tip culture medium with the stem tip facing upwards and the section facing downwards for culture, next, culturing the stem tips for 40-50 days under the conditions of a temperature within the range of 22+/-2 DEG C, the illumination time of 12h-15h/d and the illumination intensity of 2000-3000lx, in the period, transferring the stem tips above 0.5cm long to an MS culture medium, and continuously culturing the undifferentiated stem tip callus or small plantlets less than 0.5cm long, and finally, detecting the stem tip seedlings by use of enzyme linked immunosorbent assay, and remaining the satisfied stem tip seedlings, thereby obtaining the potato seedlings without PVS through the whole period of 120-180 days. The method is high in PVS virus removal rate, suitable for batch operations of removing the PVS viruses of the potato variety resources, and capable of obtaining the satisfied seedlings at a time and avoiding repeated labor; besides, the method is high in virus removal rate and stable in virus removal effect, and the detection cost can be reduced.

The invention relates to method and agent box for detecting mammary gland globulin in the field of biology chemistry. The latex fixation leptospiral method comprises the steps of: a) mixing the mammary gland globulin antibody marked emulsion particle with the tested sample, b) quoting the test result by weather it has agglutination reaction, wherein the mammary gland globulin antibody marked emulsion particle is the emulsion particle which uses mammary gland globulin antibody R028 or R048 marked emulsion particle with the dilemma 2-10um; preserving the marked emulsion particle inside the HEPES buffer solution with pH 8.2. It also discloses a method for using the anti-mammary gland globulin monoclonal antibody and skin factor mark anti-mammary gland globulin monoclonal antibody to quantity test the mammary gland globulin by double clamp heart enzyme immune adsorption method. It also discloses a RT-PCR test mammary gland globulin expressing method and provides the agent box used to test the mammary gland globulin.

The invention discloses a rapid propagation method of ginger virus-free seedlings by one step. The rapid propagation method comprises the following steps: A, selecting tender shoots germinating on ginger rhizomes as explants; B, carrying out heat treatment to the explants, so as to detoxify the explants; C, disinfecting the detoxified explants with ethyl alcohol and mercury bichloride; D, peeling stem tips by a dissecting microscope; E, inducing the stem tips to grow micro ginger cluster buds under light condition and constant temperature condition; F, carrying out virus detection on the micro ginger cluster buds by an enzyme-linked immunosorbent assay (ELISA); G, carrying out subculture multiplication culture on the virus-free micro ginger cluster buds; H, transplanting virus-free micro ginger cluster buds with roots after plant division. By the utilization of the rapid propagation method, various viruses and germs in ginger bodies are removed effectively, germ-free and virus-free ginger seedlings are cultured, virus-free healthy cultivation of the ginger is realized, space occupation of propagation is small, propagation speed is high, one virus-free bud can propagate more than hundred thousand virus-free ginger seedlings for one year, a problem of virus-free ginger seedling supplying in plantation of ginger is solved effectively, good quality of the ginger is recovered, the disease occurrence is reduced, and the output and quality of the ginger are improved.

The invention discloses a magnetic particle-based quantitative chemiluminescent assay kit for an anti-double-stranded DNA antibody IgG. The kit comprises an anti-double-stranded DNA antibody IgG calibrator, an anti-double-stranded DNA antibody IgG quality control product, a Tris buffer containing biotin-labeled double-stranded DNA antigen and bovine serum albumin, a Tris buffer containing an alkaline phosphatase-labeled goat anti-human polyclonal antibody and bovine serum albumin, a Tris buffer containing streptavidin-labeled magnetic particles and bovine serum albumin, and a rinsing solution. The detection method using the kit improves sensitivity and a linearity range by 3 to 5 orders of magnitudes on the basis of a traditional membrane strip immunization method and a traditional enzyme linked immunosorbent assay method, realizes real quantitative determination, is rapid in reaction and reliable in results, can be automatically used in cooperation with an automatic chemiluminescence immunity analyzer and is of irreplaceable important value to clinical diagnosis.

The invention discloses a method for detecting allergen-specific antibodies in serum. The method comprises the following steps: coupling anti-human IgE antibodies with carboxyl surface modified magnetic microballoons to obtain immunomagnetic microspheres; incubating the immunomagnetic microspheres with serum to be measured so as to enable the immunomagnetic microspheres to bind to IgE in the serum to be measured; carrying out magnetic separation to obtain immunomagnetic microsphere-IgE conjugates, dissolving deposition of the magnetic separation in a buffer solution, adding the mixed solution into apertures of an enzyme label plate which is coated with allergen, and carrying out detection by the ELISA adsorption method after incubation and plate washing. According to the invention, magnetic microballoons are coupled with anti-human IgE antibodies to prepare immunomagnetic microspheres which are mixed with serum to be measured for incubation and are bond to all the IgE in serum; the immunomagnetic microspheres are enriched and IgE is separated; the ELISA adsorption method is employed to detect whether there is specific IgE bond to allergen in serum. The method provided in the invention enables all the IgE to be separated from serum through immunomagnetic microspheres, impurities to be removed, and sensitivity, specificity and accuracy of ELISA adsorption detection to be improved.

The invention provides application of bacillus megaterium in degradation of peanut meal aflatoxin B1, and belongs to the technical field of microbe application. A method comprises the following steps of: sterilizing crushed target peanut meal, adding a certain amount of aflatoxin B1, then adding bacillus megaterium liquid in an amount which is 5 to 10 percent of the inoculation amount, adding sterile water in a ratio of materials to water of 1:0.5-1.5, and fermenting for 48 to 72 hours at the temperature of between 30 and 40 DEG C; and adding a solvent into the peanut meal after fermentation is completed, shaking a table for 10 to 50 minutes, extracting the residual aflatoxin B1, and detecting the content of the aflatoxin B1 by using enzyme linked immunosorbent assay. The method has the advantages of capability of effectively reducing the aflatoxin B1 content of the peanut meal, low energy consumption, low environmental pollution and no need of wastewater treatment, strict sterile operation or continuous ventilation.

The invention discloses an enzyme-linked immunosorbent assay method for detecting cadmium-metallothionein in the flesh of a shell. The method comprises the following steps of: inducing the shell to synthesize the cadmium-metallothionein by using cadmium chloride, inducing a rabbit to synthesize an antibody by using purified metallothionein, separating and purifying the antibody, using an electrophoretically pure rabbit anti-shell metallothionein antibody as a primary antibody and a goat anti-rabbit enzyme labelled antibody as a secondary antibody, and assaying the cadmium-metallothionein content of the shell by using the indirect competitive enzyme-linked immunosorbent assay method. The method has the advantages of scientific and rational principle and relatively mature technical process and is used for assaying the cadmium-metallothionein content of the flesh of the shell so as to indirectly reflect the cadmium pollution of the flesh of the shell and provide a fast and sensitive detection method for assaying the heavy metal cadmium in the flesh of the shell.

The invention provides a preparing method and application of mimic enzyme with double catalysis functions based on a hemin mediation gold mineralization path. The one-pot type in-situ synthesis method is adopted, the hemin and chloroauric acid are mixed under the alkaline condition, then gold in-situ biological mineralization is achieved with the hemin as a reducing agent and a stabilizing agent, and a Hemin-AuNCs compound with Hemin peroxidase catalysis and gold catalysis activity is prepared. The function of a nano wire of the Hemin-AuNCs compound is brought into play, the electronic transmission capability of the Hemin catalysis reaction is promoted, meanwhile, the adsorption capability for a substrate is enhanced, and the catalysis activity of the Hemin-AuNCs compound is obviously higher than that of common Hemin by four or more times. Complex photoelectric instrument usage is not involved in the preparing method, and the beneficial effects that the catalysis activity is high, operation is simple, response is fast and cost is low are achieved. The cancer biomarker alpha fetoprotein immunoassay serves as the example, the Hemin-AuNCs compound is applied to marking of an AFP antibody, then, the enzyme linked immunosorbent assay is adopted, and high-sensitivity detection on the cancer biomarker alpha fetoprotein in blood is achieved through the enzymatic catalysis and gold catalysis silver deposition signal amplification path.

The invention discloses a chlorothalonil antigen, an antibody preparation method and a residual chlorothalonil ELISA (Enzyme-Linked Lmmuno Sorbent Assay) detection method. Chlorothalonil is used as an initial reactant; an amino group is used for replacing a 4-bite chlorine atom; an artificial antigen is prepared by using a 1'1-carbonyl diimidazole method; a rabbit-resistant chlorothalonil polyclonal antibody can be obtained by an immune New Zealand white rabbit; the antiserum titer can be determined to be 1:5.12*10<4> by using the ELISA; and on the basis, the residual chlorothalonil ELISA detection method can be established; and the lowest detection concentration by using the method is 0.383ng/mL.

The invention discloses an envelope protein VP28 idiotype monoclonal antibody against shrimp white spot syndrome virus (WSSV) and a preparation method thereof. The antibody is secreted by a hybridoma cells with the collection number of CCTCC-CT200938, is prepared by taking anti-WSSV-VP28 monoclonal antibody (Ab1) as antigen, can bind with anti-WSSV-VP28 antibody of hare, and has the capability of competing with WSSV to bind with the anti-WSSV-VP28 antibody of the hare. The anti-WSSV-VP28 idiotype monoclonal antibody (Ab3) prepared by taking the antibody as antigen can bind with the WSSV, the binding site of the anti-WSSV-VP28 idiotype monoclonal antibody (Ab3) and the WSSV is located on an envelope, and the Ab3 can neutralize WSSV infection and has Ab1 properties. In the invention, the idiotype antibody is applied in the research of WSSV for the first time; a screening system is established, which uses an indirect enzyme-linked immunosorbent assay (ELISA) method and a competitive enzyme-linked immunosorbent assay (ELISA) method for detection; the fact that Ab3 has properties of Ab1 is proved by adopting an indirect immnnofluotesent method (IIF), a gold labeling immunoelectron microscopic method and crayfish in vivo neutralization tests, thus proving that the monoclonal antibody in the invention has the property to simulate original antigen WSSV-VP28.

The invention discloses a detection kit for an anti-mitochondrial antibody IgG (Immunoglobulin G). The detection kit is characterized by comprising a first reagent, a second reagent, a magnetic particle separation reagent, a chemiluminescent substrate, a calibration material, a quality control material and a cleaning solution, wherein the first reagent is a solution containing an M2 antigen coupled with biotin; part of the M2 antigen is a naturally-extracted M2 antigen, and the other part of the M2 antigen is a recombinant M2 antigen; the second reagent is a solution containing an anti-human IgG antibody coupled with alkaline phosphatase. According to the detection kit disclosed by the invention, by using the complementarity of antigens from multiple sources in the first reagent, clinicalsensitivity is greatly improved, and linear range is greatly widened; standard time of coming out the result after all the flows are finished is 50 minutes; compared with an enzyme linked immunosorbent assay, the detection kit has the advantage that reaction time is greatly shortened.

The invention relataes to novel evolutionary immunoglobulin binding molecules, as well as their preparation methods and applications. The invention discloses separated evolutionary immunoglobulin binding molecules, which are proteins with amino acid sequences as shown by SEQ ID NO : 1, or conservative variant proteins with immunoglobulin binding activity. The invention also discloses the gene encoding, genetic engineering preparation methods and applications of immunoglobulin binding molecules. The disclosed immunoglobulin molecule broad-spectrum combined with various immuneglobulin shows high immunoglobulin whole molecule binding activity, and can be used in large-scale purification of genetic engineering antibodies, purification of natural antibodies and monoclonal antibodies, enzyme-linked immunosorbent assay, and immuno-chromatography and immunohistochemical methods for immune antibody detection and diagnosis.

The invention belongs to the technical field of immunodetection and specifically relates to a quantum dot immunochromatography test strip for quickly detecting beta-amyloid protein and an application.The quantum dot immunochromatography test strip for quickly detecting beta-amyloid protein comprises nitrocellulose membranes arranged in a detection area and a quality control area, glass fiber membranes coating quantum dot micro-particles and coupled with Abeta40 and Abeta42 polyclonal antibodies, a base plate, a sample cushion and a water-absorbing cushion. The quantum dot immunochromatographytest strip for quickly detecting beta-amyloid protein provided by the invention is capable of realizing simultaneous detection of Abeta40 and Abeta42 antigen indexes through once reaction by utilizing a quantum dot immunochromatography technology, is capable of avoiding the problem of intersection of different proteins and has the advantages of short detection time, sensitivity 10 times or aboveof that of enzyme-linked immunosorbent method, high specificity, rapid response, high accuracy and quantification accuracy.

The invention relates to the technical field of immunological detection and concretely relates to a kit for magnetic particle chemiluminescent quantitative detection of an anti-SS-B antibody IgG and its preparation method and detection method. The kit for magnetic particle chemiluminescent quantitative detection of an anti-SS-B antibody IgG comprises an Anti-SS-B IgG calibration material, an anti-SS-B IgG regent 1, an anti-SS-B IgG regent 2, an anti-SS-B IgG magnetic separation regent, an anti-SS-B IgG quality control material and a cleaning liquid. The invention also discloses a preparation method and detection method of the kit. Based on the traditional membrane stripe immunization method and enzyme-linked immunosorbent assay, the detection method improves sensitivity and a linear range by 3-5 orders of magnitudes, realizes quantitative determination, has the advantages of high sensitivity, good specificity, good accuracy, low cost, simple operation and objective result determination, realizes full automation by a full-automatic chemiluminescence immunity analyzer and has a wide application prospect.

The invention relates to a serum CENPF antibody quantitative detection kit which can be used for specifically and quantitatively detecting the level of a CENPF antibody in a serum specimen. According to the serum CENPF antibody quantitative detection kit, recombinant CENPF antigen protein is coated in a 96-pore enzyme label board, the level of the CENPF antibody in the serum specimen can be quantitatively tested by using an enzyme linked immunosorbent assay, and the disease state of a liver cell cancer patient or high risk groups can be evaluated according to the level of the CENPF antibody in the serum. By adopting the serum CENPF antibody quantitative detection kit, a novel liver cell cancer screening and early diagnosis method is provided, and the sensitivity of the detected serum CENPF self-antibody is remarkably superior to that of a conventional clinical serum marker AFP when being used for diagnosing liver cell cancer.