86results about How to "Reduce non-specific reactions" 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 method of detecting the UDG activity by enzyme-mediated two-step serial signal amplification based on excision repair. The method comprises the following steps: (1) adding a UDG active substrate in an excision reaction buffer solution to carry out excision repair reaction, excising a uracil base of the UDG active substrate, and leaving an abasic site; and (2) adding an excision repair reaction product in an amplification reaction buffer solution to carry out enzyme-assisted two-step serial signal amplification reaction, namely inducing strand displacement reaction and index amplification reaction, generating strengthened fluorescence signals in a circulation manner, and determining the UDG activity through weakness of the fluorescence signals. According to the method provided by the invention, ultrahigh sensitive detection to the activity of uracil DNA glycosylase (UDG) is realized by utilizing high amplification efficiency of constant-temperature index amplification reaction and specific circulation digestion of ribonuclease H.

The invention relates to a kit for detecting Southeast Asia type alpha-thalassemia and discloses a sample treating fluid of the kit. The kit is characterized in that in the sample treating fluid, the content W/V of ammonium chloride is 0.83%, the required content W/V of casein ranges from 0.2% to 0.8% and the content V/V of glycerin ranges from 4% to 6%. The kit can conveniently and effectively detect Zeta globin chains and assist in diagnosing the Southeast Asia type alpha-thalassemia.

The present invention relates to a rapid, efficient and accurate bovine Brucella colloidal gold diagnostic method. According to the present invention, Brucella strain S2 lipopolysaccharide (LPS) is adopted as a testing line coating antigen so as to improve the detection sensitivity; the LPS purification and purity determination method is improved, such that the developed kit has good specificity; the monoclonal antibody on the Fc terminal of the mouse anti-bovine IgG is used to label the colloidal gold to prepare the colloidal gold pad of the colloidal gold antibody detection test paper strip so as to further improve the detection specificity; the bovine Brucella colloidal gold antibody detection test paper strip can be used for detecting the Brucella antibody in the bovine serum, and has characteristics of strong specificity, high sensitivity, convenience, rapidness, and the like; and the complex detection equipment is not required for the Brucella detection, and the bovine Brucella colloidal gold antibody detection test paper strip is especially suitable for cattle farm breeding staffs and grassroots veterinarians.

The invention provides a novel severe acute respiratory syndrome coronavirus 2 N proteantigen variant and an application thereof to detection of the novel severe acute respiratory syndrome coronavirus2 antibody, and relates to variants of N protein important epitope polypeptide. The variants reduce the non-specific reaction of the novel severe acute respiratory syndrome coronavirus 2 protein as acapture antigen with other coronavirus antibodies such as SARS and human influenza coronavirus antibodies. The polypeptide has coronavirus N protein epitope peptide activity, and cysteine residues are added to the C end of the polypeptide. Amino acid residue sites of the mutation are selected from: lysine at the 342rd site, lysine at the 347 site, lysine at the 387 site and lysine atthe 388 site are mutated into amino acids with relatively low homology. The sequence of the N protein 340-419 polypeptide is shown as SEQ ID NO 7. The invention further discloses an antigen composition, use and method of the variant.

The invention discloses an improved experimental buffer solution. The improved experimental buffer solution comprises a basic experimental buffer solution, sodium chloride and a dissociation agent, wherein the dissociation agent is at least one selected from the group consisting of 8-anilinonaphthalene-1-sulfonate, thimerosal, sodium salicylate and sodium trichloroacetate. The invention also discloses the application of the improved experimental buffer solution. The improved experimental buffer solution provided by the invention can effectively reduce electrostatic force, promote the dissolution and dissociation of non-specific immunoreactive or non-specifically adsorbed low-affinity protein ligands, and reduce the non-specific binding of antigens and antibodies; and the improved experimental buffer solution can promote the dissociation of non-specific immunoreactive or non-specifically adsorbed protein ligands under the action of the dissociation agent, so non-specific reactions of immunoassay are reduced, and detection resolution, accuracy and precision are improved.

The invention provides a novel HIV recombined multi-epitope fusion antigen and a detection kit for preparing through the antigen. Flexible structured genes are added into the main epitope genes of HIV-1 gp41, M group and O group, the main epitope genes of HIV-1gp120 and the main epitope genes of HIV-2gp36, so that all the gene DNA are linked to be an artificial gene sequence. The sequence is cloned to an expression vector for ectopic expression, so as to establish immunocompetent high-expression cell strain, finally the sequence is fermented and induced into efficient expression, and chromatographic separation and purification is performed on the multi-antigen epitope fusion antigen of the HIV. The antigen is applied to the immune detection reagent, as the unnecessary gene sequence is reduced through the fusion antigen, the non-specific action is reduced for more than 50%, and the synergistic effect of the multi-antigen epitope of the same antigen increases the detection sensitivity to be 2 to 3 times.

The invention relates to an enzyme-linked immunosorbent assay (ELISA) kit and a preparation method thereof, and more specifically relates to a novel anti-human FN1 protein monoclonal antibody coated ELISA plate and a preparation method of the ELISA kit. A key step of the preparation method is that: the specific mouse anti-human FN1 protein monoclonal antibody is prepared by genetic engineering method, and is purified; and then the ELISA plate is coated with the specific mouse anti-human FN1 protein monoclonal antibody, and is used as a constituent part of the ELISA kit. The ELISA kit also comprises FN1 recombined protein standard, polyclonal antibody of anti-human FN1, horse radish peroxidase-labeled second antibody, a sample diluent, a washing liquid, an antibody diluent, a colour reagent, and a stop solution. The ELISA kit can be used for rapid and convenient detection of FN1 concentration of human serum and blood plasma, and is mainly provided for scientific research or clinical research of cancer related diseases.

The invention provides a method for preparing a latex immunonephelometric assay kit with more specific performance, a more intact antibody stereostructure, higher affinity and fewer interference factors by orientedly immobilizing a single-chain antibody on the surface of a polystyrene microsphere through fusion expression of scFV and polystyrene binding peptide on the basis of the characteristic that the polystyrene binding peptide can specifically adhere on the surface of the polystyrene microsphere.

The invention relates to a blocking ELISA antibody detection kit for porcinecircovirus type 2 and a preparation method of the blocking ELISA antibody detection kit. The blocking ELISA antibody detection kit for the porcinecircovirus type 2 comprises a coated plate, a PVC2 enzyme labelled antibody, sample diluent, a concentrated washing solution (10*), a positive serum contrast, a negative serum contrast, TMB substrate diluent and a stop solution, wherein the coated plate is prepared from a purified porcine circovirus type 2 nucleocapsid protein CapC protein serving as a non-antigen, and the PVC2 enzyme labelled antibody is prepared from monoclonal antibody hybridoma which is labeled with horse radish peroxidase and secretes PCV2. According to the preparation method, the CapC protein is expressed by virtue of a prokaryotic expression system, and the blocking ELISA is established by taking a monoclonal antibody of PCV2 as the enzyme labelled antibody; and the blocking ELISA antibody detection kit has the characteristics of high specificity and sensitivity and can be applied to the detection of PCV2 antibodies of a porcine serum sample.

Disclosed is a chemiluminescence immunoassay test kit which is used to detect rubella IgG antibody, the test kit includes: positive and negative reference substance, solid-phase carrier which is coated by rubella virus antigen, sample diluent, antihuman IgG monoclonal antibody labeled by alkaline phosphatase, chemiluminescence zymolyte and condensed washing liquid. The preparing method of the test kit includes steps as following: 1) the positive and negative reference substance is prepared; 2) the solid-phase carrier is coated; 3) the sample diluent is prepared; 4) the antihuman IgG monoclonal antibody is labeled by the alkaline phosphatase; 5) the chemiluminescence zymolyte is prepared; 6) the condensed washing liquid is prepared; 7) the positive and negative reference substance, the sample diluent, the alkaline phosphatase labeled compound, the chemiluminescence zymolyte and the condensed washing liquid can be filled and packed through separate packing; 8) finished product can be made after assembly. Adopting the test kit and the preparing method, the rubella IgG antibody in the human blood serum can be detected simply, quickly, sensitively and accurately, thereby being convenient for popularization and application.

A novel human cytomegalovirus fusion protein used for detecting human cytomegalovirus antibody or antigen and preparing monoclonal or polyclonal antibody is prepared by recombining human cytomegalvirus PP150 protein with gp52 protein in such manner that the 25 amino acids at c terminal of pp150 and the 164 amino acids at C terminal of gp52 are serially linked via 2 amino acids Gly and Ser.

The invention relates to a reaction cup, and concretely discloses a reaction cup used for automatic chemical luminescence immunoassay instrument, the reaction cup comprises a cup body filled with a carrier, the profile of the cup body is in a conical shape with a gradually downward reduced diameter, an annular rectangular bench is arranged outside the cup rim of the cup body, at least three uniformly distributed circular-arc contact points are arranged at lower end of the annular rectangular bench; the bottom of a cup core space is an arc-shaped spherical surface, the upper part of the cup core space is a conical surface cooperated to the profile of the cup body, smooth transitioning is realized between the arc-shaped spherical surface and the conical surface; the reaction cup can employ a staked mode for storage; wherein the annular rectangular bench can clamp the reaction card in a reaction aperture of an incubation apparatus, at least three uniformly distributed circular-arc contact points are arranged at lower end of the annular rectangular bench, when the reaction cups are stakes, the contact area is little, so that the adhesion probability between the reaction cups while loading can be reduced.

The invention relates to a preparation process for a hybridoma secreting a monoclonal antibody repelling human asparaginate hydroxylase. The method comprises the following steps: taking the recombinant enkaryon expression plasmid DNA of a human asparaginate hydroxylase HAAH coded gene, injecting the DNA into the skeletal muscle of a white rat for DNA immunity, carrying out human asparaginate hydroxylase HAAH recombinant protein immunity after 1 to 4 weeks, taking the splenic cell and myeloma cell of the immune white rat for cell fusion after 2 to 5 days, carrying out antibody detection, and cloning the positive cells in the antibody detection till obtaining the hybridoma secreting the monoclonal antibody repelling human asparaginate hydroxylase. The invention solves the technical problems of bad specificity, long immune cycle and low efficiency in the prior art. The hybridoma has clear immunogenicity, high purity, few non-specific reactions, and low use level of antigen with a total immune cycle of only 6 to 8 weeks.

The invention discloses a monoclonal antibody 7D7 of nonstructural protein of a foot-and-mouth disease virus (FMDV), a foot-and-mouth disease virus nonstructural protein antibody detection kit containing the monoclonal antibody and application of the monoclonal antibody. The antibody detection kit provided by the invention has high sensitivity and a linear range of detection can be remarkably widened; in a detection process, the monoclonal antibody is not limited by animal species and is applicable to large-scale screening of foot-and-mouth diseases of cloven hoof type animals. A vaccine prepared from a conjugate containing the monoclonal antibody 7D7 can be used for removing the nonstructural protein; after the animals are immunized, interference to the nonstructural protein antibody of the FMDV can be avoided, so that infected animals and immunized animals are accurately distinguished.

The invention discloses a glucagon detection kit. The glucagon detection kit comprises a glucagon-antibody-coated solid-phase carrier (1), a horse-radish-peroxidase-labeled glucagon detection antibody (2), an enzyme-labeled antibody diluent (3), a glucagon calibrator (4), a washing liquid (5) and an enzyme substrate solution (6), wherein the glucagon antibody can specifically identify an N-terminal monoclonal antibody of human glucagon; and the glucagon detection antibody can specifically identify a C-terminal monoclonal antibody of human glucagon. The invention also discloses a detection method of the glucagon detection kit. The kit is simple in formula and convenient to use, and can sensitively, quickly and accurately measure glucagons in both blood and plasma at the room temperature in clinical test.

The invention relates to a reagent kit for assessing endometrial receptivity. The reagent kit comprises gDNA scavenger buffer liquid, a gDNA scavenger, cDNA reverse transcription buffer liquid, a cDNAreverse transcription mixture, a reverse transcription primer mixture, real-time fluorescent quantitation polymerase chain reaction liquid, a probe, a positive control standard product and enzyme-free water. The probe is combined with a target gene through molecular hybridization to generate hybridized signals, so that the target gene in massive genomes can be shown. A specific sequence of reverse transcription products is combined with detection of probe signals, so that expression level of specific genes can be detected, and the endometrial receptivity can be conjectured. The manner is simple to operate, high in sensitivity, high in specificity, accurate in detection results, objective and quick, so that a clinician can be helped to clear the state of the endometrial receptivity, and the reagent kit can provide a basis for early diagnosis, target intervention and guidance of an embryo transplanting opportunity for infertility patients.

The invention provides a blocking method for activated quantum dots. The activated quantum dots are blocked by adopting polymer inert protein formed by cross-linking reaction of 1-ethyl-3-3-(3-dimethyl aminopropyl)-carbodiimide (EDC) and inert protein. The method disclosed by the invention has the advantages that effective blocking of the surfaces of the activated quantum dots can be ensured and protein linking in the blocking process is firmer, and thereby the sensitivity and the stability of a reagent are improved and basic performance of the quantum dot immunochromatography reagent is improved. The invention also aims at providing application of the method in an immunofluorescence assay kit for the quantum dots.