17 results about "Brucella antibody" patented technology

The invention is applicable to the technical field of biology, and provides an L7 / L12-PADRE sequence-multi-linked B cell epitope recombinant protein and a kit for detecting an anti-Brucella antibody. The recombinant protein is a multi-epitope tandem recombinant protein composed of Brucella ribosome L7 / L12 protein, a PADRE polypeptide sequence and multiple B cell epitopes, the amino acid sequence of the recombinant protein is as shown in SEQ ID No.1, and the recombinant protein is good in antigenicity. The anti-brucella antibody indirect ELISA (iELISA) detection method and kit established by taking the recombinant protein as the coating antigen have the characteristics of high sensitivity, strong specificity and good repeatability, are suitable for detecting the condition of generating the anti-brucella antibody by an organism, can clarify the immune background of brucella infection or brucellosis vaccine, and can be used for detecting the brucella infection or brucellosis vaccine. Basic data is provided for prevention and control of the Brucella disease of humans and animals, and meanwhile, the kit is conveniently applied to large-scale sample detection and epidemiological monitoring.

The application belongs to the technical field of biological pharmacy, and particularly relates to a bovine-derived mouse anti-Brucella antibody and a preparation method and application thereof. A bovine-derived mouse anti-Brucella antibody is prepared by synthesizing a tandem epitope protein of Brucella OMP25 proteins of bovine species, sheep testis species and canine species, immunizing mice, preparing a monoclonal antibody, cloning CDR fragments in Hv and Lv regions of the antibody to corresponding positions of bovine-derived IgG, and then performing overall mutation modification on the recombinant IgG to perform bovine-derived modification. The bovine-derived mouse anti-Brucella antibody preparation has good immunogenicity and protection effect, and can block and inhibit the propagation and spread of Brucella of bovine species, canine species and sheep testis species in an animal body.

The invention relates to the technical field of biological detection, and particularly discloses a preparation method and application of a Brucella antibody immunochromatography detection test strip based on colloidal carbon. The Brucella antibody immunochromatography detection test strip provided by the invention comprises a PVC (Polyvinyl Chloride) bottom plate, a sample pad, a combination pad, a nitrocellulose membrane and a water absorption pad are arranged on the PVC bottom plate; the binding pad contains lipopolysaccharide of colloidal carbon labeled brucella, the nitrocellulose membrane is provided with brucella lipopolysaccharide and a goat anti-mouse igG antibody which are respectively used as a detection line (T) and a quality control line (C), and the colloidal carbon technology-based immunochromatography test paper is established by detecting different specimens and comparing with other detection methods. The test strip prepared by the invention has the advantages of no subjective influence of a detector, simplicity and convenience in operation, high detection sensitivity, strong specificity and the like.

The invention provides a specific polypeptide of Brucella, the amino acid sequence of the specific polypeptide is as shown in SEQ ID No.1, and the specific polypeptide is derived from BMEI1871 protein of Brucella. The invention also provides a recombinant vector or an expression cassette for expressing the specific polypeptide, and a recombinant cell or a recombinant bacterium. The invention verifies the application of the specific polypeptide in preparation of drugs or reagents for preventing, detecting or diagnosing brucellosis, and also provides an ELISA (enzyme-linked immuno sorbent assay) detection kit for detecting brucellosis. According to the present invention, the specific polypeptide of the brucella BMEI1871 protein is adopted as the antigen to establish the ELISA method for detecting the brucella antibody, such that the accuracy is high, and the powerful support is provided for brucellosis purification. The antigen source is convenient, the stability is high, the repeatability is strong, and the defects of using whole bacteria or lipopolysaccharide as the antigen are overcome.

The invention provides an antibody combination and application thereof in a test strip for detecting IL-1Ra, IL-1beta and Brucella antibodies, and belongs to the technical field of cell factor and Brucella detection. The antibody combination provided by the invention comprises an IL-1beta paired antibody and an IL-1Ra paired antibody, the IL-1beta paired antibody comprises an IL-1beta-3-3 antibody and an IL-1beta-1S-4 antibody, and the IL-1beta paired antibody comprises an IL-1beta-3-3 antibody The IL-1Ra paired antibody comprises an IL-1Ra-2S-2 antibody and an IL-1Ra-1S-23 antibody, and the IL-1Ra paired antibody can be specifically combined with an IL-1beta recombinant protein, an IL-1Ra recombinant protein, and IL-1beta and IL-1Ra in natural sheep serum. The invention further provides a fluorescence immunochromatography test strip which can be used for qualitative and quantitative detection at the same time and can be used for detection of IL-1Ra, IL-1beta and Brucella antibodies in serum.

This invention discloses a Brucella lipopolysaccharide monoclonal antibody, the heavy chain variable region sequence of which is shown in SEQ ID NO.1, and the light chain variable region sequence is shown in SEQ ID NO.2. This invention also discloses a kit for detecting Brucella antibodies, the kit comprising an effective amount of monoclonal antibody and Brucella lipopolysaccharide, as well as matching detection reagents. The Brucella antibody detection kit prepared by this invention has good specificity and sensitivity, and is suitable for field applications of Brucella.

The application belongs to the technical field of biotechnology, and provides a L7 / L12-PADRE sequence-multiple B cell epitope recombinant protein and kit for detecting anti-Brucella antibody. The recombinant protein is a multi-epitope tandem recombinant protein composed of Brucella ribosome L7 / L12 protein, PADRE polypeptide sequence and multiple B cell epitopes, and the amino acid sequence of the recombinant protein is shown as SEQ ID No. 1, and the antigenicity is good. The anti-Brucella antibody indirect ELISA (iELISA) detection method and kit established by taking the recombinant protein as a coating antigen have the characteristics of high sensitivity, strong specificity and good repeatability, are suitable for detecting the anti-Brucella antibody produced by the organism, can clearly determine the Brucella infection or Brucella vaccine immunization background, provide basic data for preventing and controlling the human-animal shared Brucella disease, and are simultaneously convenient for being applied to large-scale sample detection and epidemiological monitoring.

The application relates to the technical field of biological detection, and particularly discloses a bovine brucella antibody immunochromatographic detection test strip as well as a preparation method and application thereof. The bovine brucella antibody immunochromatographic detection test strip provided by the application comprises a PVC bottom plate, a sample pad, a combination pad, a nitrocellulose membrane and a water absorption pad are arranged on the PVC bottom plate, the combination pad contains quantum dot microsphere labeled mouse anti-bovine IgG monoclonal antibody, the nitrocellulose membrane is provided with a detection T line and a quality control C line, the detection T line is coated with a brucella LPS antigen, and the quality control C line is coated with a goat anti-mouse IgG polyclonal antibody. The bovine brucella antibody immunochromatographic detection test strip provided by the application has the advantages of being not influenced by subjective factors of a tester, simple and convenient operation, high detection sensitivity and specificity and the like.

The invention discloses an agglutination-labeled complement C1q binding reaction (A-LCFA) detection technical method based on fusion agglutination reaction and labeled complement C1q binding reaction and application of the A-LCFA detection technical method. According to the method, high simplicity and convenience of the agglutination reaction are combined with high specificity of the complement C1q binding reaction and high sensitivity of a labeling technology; the efficient detection of the target antibody or antigen is realized. The method comprises the following specific steps: reacting a granular antigen or antibody with a sample to be detected to form an agglutination complex, adding a complement C1q binding antigen-antibody complex, and carrying out signal detection by marking a complement C1q antibody. The method is suitable for the field of immunological antibody or antigen detection, and has the characteristics of simplicity and convenience in operation, high specificity and high sensitivity. The embodiment shows successful application of the Brucella antibody detection kit in Brucella antibody detection, and the Brucella antibody detection kit comprises a plate agglutination mode and a column agglutination mode. The technology provides a new solution for immunological detection, and has remarkable practical value and popularization potential.

The invention discloses a rapid Brucella antibody detection kit, belongs to the technical field of antibody detection, and aims to solve the problem of detection misjudgment caused by the fact that a pretreated blood sample is not easy to quantitatively drop into a reagent card in the kit, the rapid Brucella antibody detection kit comprises a kit main body, and the top of the kit main body is rotatably connected with a box cover; handles are fixedly connected to the two sides of the kit body, a plurality of insertion holes are formed in one side of the kit body in a penetrating mode, a limiting assembly is arranged in the kit body, a plurality of first long grooves are formed in the limiting assembly in a penetrating mode, a detection assembly is slidably connected into the limiting assembly, and the detection assembly communicates with the first long grooves. A plurality of positioning assemblies are uniformly arranged on one side of the kit main body, the positioning assemblies are attached to the detection assembly, and a lifting assembly is arranged on one side of the limiting assembly, so that the sample adding amount of multiple channels is consistent, quantitative liquid dropping is performed, and the sample adding and detection precision is improved.

The invention is applicable to the technical field of biology, and provides a nano antibody for resisting Brucella Omp25 protein and application of the nano antibody in competitive ELISA (enzyme-linked immuno sorbent assay) detection. The amino acid sequence of the nano antibody is as shown in SEQ ID NO. 3. Omp25 protein of prokaryotic expression brucella is used as an immunogen to immunize alpaca, peripheral blood lymphocyte total RNA of the alpaca is extracted, a VHH fragment is amplified through reverse transcription and two rounds of nested PCR, the VHH fragment and a pComb3Xss vector are subjected to enzyme digestion and connection and transformed into a TG1 competent cell, a phage display technology is utilized to perform multiple rounds of panning, and the recombinant protein is obtained. And screening out the nano antibody for resisting the Brucella Omp25 protein. The competitive ELISA detection method established by using the nano antibody can effectively detect the level of the Brucella antibody in sheep serum, and is low in detection limit and high in sensitivity; the developed matched kit is high in specificity, and the detection time is greatly shortened.

The application belongs to the field of biotechnology, and provides a nano antibody against Brucella Omp25 protein and application thereof in competitive ELISA detection. The amino acid sequence of the nano antibody is shown as SEQ ID NO. 3. The application immunizes a llama with prokaryotic expressed Brucella Omp25 protein as an immunogen, extracts total RNA of peripheral blood lymphocytes, performs reverse transcription and two rounds of nested PCR to amplify a VHH fragment, performs enzyme digestion and connection on the VHH fragment and a pComb3Xss vector, and transforms the VHH fragment into TG1 competent cells. Through multiple rounds of panning by using phage display technology, the nano antibody against Brucella Omp25 protein is screened. The competitive ELISA detection method established by using the nano antibody can effectively detect the Brucella antibody level in sheep serum, has low detection limit and high sensitivity; and the developed matching kit has high specificity and greatly shortens the detection time.

The invention discloses a fluorescent microsphere immunochromatography test strip for detecting a Brucella antibody and application of the fluorescent microsphere immunochromatography test strip. On the basis of systematic bioinformatics analysis of multiple dimensions such as epitope coverage breadth, immunogenicity intensity and antigen accessibility, OMP22, VirB12, BP26 and OMP10 are selected to form a detection antigen combination, the antigen combination is used as a detection line of a fluorescent microsphere immunochromatography test strip, goat anti-rabbit IgG with the coating concentration of 1.0 mg / mL is used as a quality control line, fluorescent microspheres are used as a marker, and the fluorescent microsphere immunochromatography test strip can be used for detecting the immunogenicity of the fluorescent microsphere immunochromatography test strip. The fluorescent microsphere immunochromatography test strip for detecting the Brucella antibody is developed by coupling staphylococcus aureus A protein and spraying on a glass cellulose membrane as a combination pad. Experiments prove that the test strip has good sensitivity and specificity, and the coincidence rate of the test strip and a test tube agglutination test (SAT) detection result reaches 90%.

The invention relates to the technical field of biology, in particular to a kit for detecting a livestock brucella antibody and application of the kit. The magnetic particle chemiluminescence method comprises the following steps: coating magnetic beads, a signal tracer agent, a reaction diluent and the like; the coated magnetic beads are magnetic beads coated with Brucella specific lipopolysaccharide antigens, and the dilution concentration of the coated magnetic beads is 0.05-0.15 mg / mL; the signal tracer agent is recombinant streptococcus protein G marked by acridinium ester, and the dilution ratio is 1: (8000-12000); the reaction diluent is a phosphate buffer, the concentration of the phosphate buffer is 0.01-0.03 M, and the pH value of the phosphate buffer is 7.3-7.5. The detection method provided by the invention has high specificity, sensitivity, repeatability and reliability, and can be used for the detection purposes of brucellosis epidemiological investigation, immune antibody evaluation and the like.

The application discloses a kind of detection anti-Brucella antibody multi-link B cell epitope recombinant protein and its application, it belongs to the field of biotechnology and immunodiagnosis technique, wherein, multi-link B cell epitope recombinant protein is formed by the B cell epitope of seven key outer membrane proteins Omp10, Omp16, Omp19, Omp25, Omp31, Omp2b, BP26 of Brucella, its amino acid sequence is as shown in sequence table SEQ ID NO:1.The multi-link B cell epitope recombinant protein has good antigenicity and reactivity, and the deer serum indirect ELISA detection system established with it as coating antigen shows excellent detection performance, has high sensitivity, strong specificity and good repeatability, and can accurately detect the level of anti-Brucella antibody in deer serum sample.

This invention discloses a method for preparing Brucella antibodies, relating to the field of Brucella antibody preparation technology. Based on one or more combinations of Brucella outer membrane proteins Omp10, Omp19, and BP26, a recombinant antigen is prepared, and the prokaryotic expression codon is optimized. Alpacas are immunized subcutaneously with the recombinant antigen and adjuvant at multiple sites, with booster injections every two weeks for three weeks, to isolate highly active lymphocytes. Total RNA is extracted from the lymphocytes, reverse transcribed to synthesize cDNA, and the VHH gene is specifically amplified by nested PCR to accurately obtain the antibody coding sequence. The VHH gene is cloned into a phage vector, and a high-capacity display library is constructed by electroporation. Solid-phase panning is performed using the recombinant antigen as a target, followed by sequencing to construct a prokaryotic expression vector. Recombinant nanobodies are induced to express, purified by affinity chromatography, and elution conditions are optimized to obtain high-purity, high-yield target proteins. The optimized prokaryotic expression and purification process results in nanobodies with high soluble expression rate, high purification yield, and stable activity, achieving large-scale, low-cost preparation.

The application belongs to the technical field of animal epidemic disease immunity detection, and particularly relates to a Brucella antibody detection test paper and application, which comprises a bottom plate, a chromatography membrane arranged on the bottom plate, one end of the chromatography membrane is provided with a sample processing layer, and the chromatography membrane is further provided with a quality control line and at least one detection line; the sample processing layer comprises chicken IgY marked latex microspheres, Protein G protein marked latex microspheres or / and Fcα / μR receptor marked latex microspheres, and the detection line comprises Brucella LPS; or the sample processing layer comprises Brucella LPS marked latex microspheres and chicken IgY marked latex microspheres, and the detection line comprises Protein G protein and Fcα / μR receptor; the application significantly improves the sensitivity of early diagnosis or infection period discrimination, and is good in specificity, repeatability and accuracy.