613 results about "Indirect elisa" patented technology

The invention discloses an indirect ELISA diagnosing reagent box for pig II type circular virus (PCV2) antibody. There arranged with antibody detecting board in the reagent box, enzyme combination liquid, positive comparison, negative comparison, sample diluter, 10X condensed detergent, developing liquid A and B, and stopping liquid, the detecting board of the reagent box is removable 96-apertures enzyme labeling board enclosed with PCV2 reconstructed enucleation localizing signal capsid protein (dCap), the enzyme combination liquid is goat-anti-pig IgG multi-clone antibody labeled with HRP, the positive comparison is pig PCV 2 standard positive serum, the negative comparison is pig standard negative serum. The sensitivity of the invention is high, simple, and easy to be wide applied.

The invention relates to an ELISA kit for distinctively detecting antibodies of classical swine fever (CSF) vaccine immunity and wild virus infection and a preparation method thereof. An indirect ELISA kit or a blockage ELISA kit is formed by expressing and purifying classical swine fever virus (CSFV) non-structural protein NS3, coating a solid-phase carrier and assembling with other matched reagents. The kit has the characteristic of distinctively detecting different antibodies generated by the CSF vaccine immunity and the wild virus infection. The kit can distinctively diagnose the CSF vaccine immunity and the wild virus infection.

The method comprises: 1) the GoldMag nano-particles are mixed with the double strand DNA; under certain condition, the double strand DNA is fixed on the surface of GoldMag nano-particles, or the GoldMag nano-particles are used to extract the DNA from the human blood, saliva and semen and to fix the extracted DNA on the surface of GoldMag nano-particles; 2) using indirect ELISA method, immumofluorescence method or chemiluminescent method to detect the anti double strand DNA antibody existed in the sampler under test.

The invention discloses a preparation method for a PCV-II Cap protein monoclonal antibody, an antibody and application. The invention adopts ultracentrifuged and purified PCV-II as an immunogen to immunize a BALB/c mouse by the conventional method, takes spleen cells of the immunized BALB/c mouse to fuse with SP2/0 cells, obtains two strains of hybridoma cells secreting the PCV2-Cap protein monoclonal antibodies by indirect ELISA screening, respectively names the two strains of hybridoma cells as 8-60 and 10-48, identifies biological characteristics of the two strains 8-60 and 10-48, and usesthe two strains 8-60 and 10-48 as the first antibodies to establish an indirect immunofluorescence diagnostic method. The result of the indirect immunofluorescence diagnostic method is basically consistent with that of the PCR diagnostic method, and the positive and negative coincidence rates are respectively 93.75 percent and 100 percent so as to provide reference for preventing and treating theporcine circovirus disease.

The invention belongs to the field of biological detection reagents and relates to a kit for indirect ELISA (Enzyme-Linked Immunosorbent Assay) detection of IgG or IgA antibodies of PRRSV (Porcine reproductive and respiratory syndrome virus) of pigs. The kit is used for indirect ELISA detection of the IgG or IgA antibodies of the PRRSV in serum or saliva of pigs and comprises (a) an antibody detection board, (b) an HRP-labeled goat anti-pig IgG antibody solution or HRP-labeled goat anti-pig IgA, (c) a positive control, namely PRRSV standard positive serum or saliva, (d) a negative control, namely PRRSV negative serum or saliva, (e) sample diluting liquid, (f) 10* concentrated washing liquid, (g) substrate developing liquid, and (h) stopping liquid. The kit is good in PRRSV specificity and sensitivity when being used for detection of PRRSV IgG or IgA antibodies in serum or saliva of pigs. A repeatability test also shows that the kit is good in repeatability.

The invention discloses an indirect ELISA (enzyme linked immunosorbent assay) kit for detecting a porcine epidemic diarrhea virus antibody. The indirect ELISA kit comprises a coated ELISA plate, negative control serum, positive control serum, ELISA secondary antibody, a concentrated cleaning solution, a sample diluent, a developing liquid and a stop buffer, wherein the coated ELISA plate utilizes recombinant protein Nh as a coating antigen. Research and practice show that the indirect ELISA kit has the characteristics of high specificity and sensitiveness, simplicity in operation and the like, is easy to popularize and use in a large range, has a broad market prospect, and can be used for the fields of serological investigation, antibody monitoring and vaccine immunity effect evaluation and the like of epidemic diarrhea infection condition.

The invention relates to the diagnostic medicine of animals, especially relates to a mycoplasma bovis diagnosis technology, and concretely relates to a multi-epitope fusion antigen having an amino acid sequence represented by SEQ ID NO:1 or SEQ ID NO:2, and its application in the preparation of a mycoplasma bovis diagnosis reagent. The diagnosis reagent can be used as a solid phase vector coating antigen of an indirect ELISA kit and is combined with its specific antibody, a horseradish peroxidase coupled anti-cattle IgG antibody is added and incubated, and a color development reaction is carried out, and the color development degree is proportional to the amount of the anti-mycoplasma bovis antibody in a sample to be measured. The technology has the advantages of simple operation, no need of complex equipment, low technical requirements on the laboratorial conditions and experiment personals, low detection cost, and suitableness for the large-scale development in the basic level and the culture farm; the multi-epitope fusion antigen has a low making cost and is suitable for large-scale application; and has the advantages of high sensitivity and specificity, small batch difference, and high detection result consistence because of the adoption of multi-epitope as a target.

The invention discloses an ELISA (Enzyme-Linked Immunosorbent Assay) antibody detection kit for distinguishing and diagnosing capripox field virus infection. The indirect ELISA antibody detection kit for distinguishing and diagnosing capripox field virus infection, disclosed by the invention, is internally provided with a coated antigen ELISA antibody detection board and an ELIAS secondary antibody, wherein the coated antigen is a mixture of a recombination capripox virus ORF95 protein and an ORF103 protein. The recombination proteins of ORF95 and ORF103, adopted by the invention, are convenient to prepare and purify in large quantities. The kit disclosed by the invention has a strong specificity, can effectively exclude the interference of the capripox attenuated-vaccine immunity, and specifically detects the capripox field virus infection.

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.

Indirect ELISA of deer's brucellosis belongs to the veterinary field and is used for diagnosing deer's brucellosis to improve the diagnosis level of the prior art. Biomaterials used in the assay of the invention comprise pig brucella vaccine 2 (S2) as an antigen, an enzyme labeled antibody rabbit anti-deer IgG-HRP, a standard positive serum, a standard negative serum and a serum to be assayed; and the assay comprises the following steps: coating the antigen, washing, adding the serum, adding the rabbit anti-deer IgG-HRP, adding a substrate liquid, terminating a reaction, testing by an enzyme-linked detector, adjusting to zero at 490nm, testing the OD value of each hole, and determining the result is positive when the OD value is equal to or greater than 0.32. According to the invention, the S2 is used as the antigen, the rabbit anti-deer IgG-HRP is used as the enzyme labeled antibody, and the indirect ELISA is used to diagnose the deer's brucellosis, so compared with previous diagnosis methods, the indirect ELISA of the invention has the advantages of good specificity, sensitivity and repeatability, and accurate diagnosis.

The invention discloses an indirect ELISA kit for detecting a serum type 4 aviadenovirus antibody and a detection method of the kit. The ELISA kit comprises an ELISA reaction plate, recombinant FAdV-4 fiber-2 protein, a horseradish peroxidase conjugated rabbit anti-chicken IgG antibody, coating liquid, confining liquid, a cleaning solution, a substrate developing solution and a stop buffer. The kit and detection method disclosed by the invention are applicable to rapid detection of serum type 4 aviadenovirus and have the characteristics of simplicity in operation, high specificity, high sensitivity and low cost.

The invention relates to a bovine brucella indirect ELISA antibody detection kit. The kit uses the lipopolysaccharide (LPS) of brucella suis S2 acclimated and induced by the application institution in the 1960s as the coating antigen, and improves the sensitivity of detection. The study improves the LPS purification and purity determination method, and endows the developed kit with good specificity. The kit not only can effectively eliminate the interference of conventional Gram negative bacteria to Brucella, but also can get rid of the technical defect that general brucellosis indirect ELISA (enzyme-linked immunosorbent assay) kits cannot distinguish enterocolitis Yersinia O9 and Escherichia coli O157 interference, and improves the specificity of detection. Through improvement and optimization of the main reagent formula, the reaction background of the kit is effectively controlled, and also the time required by detection is shortened.

The invention discloses a HCMVPP65 antigenemia indirect immune fluorescence method detection kit, which uses cytomegalovirus-AD169 virus strain pp65 protein as the immunogen to immune a Balb/c mouse. The spleen cells of the immunized mouse and the myeloma cells of the mouse which belongs to the same type with the immune mouse are conventionally integrated, by indirect ELISA screening and finite dilution cloning, the hybridoma cell lines of the mouse cytomegalovirus pp65 protein cloning antibody are obtained, and the characteristics of the hybridoma cell lines are identified by ELISA, immune fluorescence experiment and other methods; two monoclonal antibodies that stably secrete the pp65 protein are established successfully and named respectively as 1A6 and 4A8. A monoclonal antibody which differs from the former report and aims at the pp65 protein of the cytomegalovirus (CMV) is prepared and a method used for preparing erythrocyte fast pyrolysis is established; compared with other detection kits which belongs to the same kind, the detection kit of the invention is faster, simpler and more convenient and has higher specificity and sensitivity.

The invention discloses a chloramphenicol universal monoclonal antibody hybridoma cell strain and application thereof, belonging to the technical field of food safety immunological detection. The preparation method comprises the following steps: by taking a thiamphenicol analogue 1-(4-aminophenyl)-2-dichlorethamin-1,3-propylene glycol as a hapten, allowing the hapten to pass through diazotization coupling bovine serum albumin (BSA), thereby obtaining a complete antigen; uniformly mixing with a Freund's adjuvant, and performing subcutaneous injection immunotherapy on BALB/c mice; synthesizing an envelope antigen to be used for screening mice serum and cell supernatant by using diazotization; and fusing splenocyte of immune mice with mouse myeloma cells through a PEG method, performing indirect ELISA and indirect competitive ELISA screening and subcloning for three times, thereby obtaining a hybridoma cell strain with mass selection property. The cell strain provided by the invention has high inhibition properties on thiamphenicol, chloramphenicol and florfenicol, and the requirements on chloramphenicol stumpy immunodetection products in the current market can be met.

The invention discloses an enzyme-linked immunosorbent assay method of paraoxon-methyl, belonging to the technical field of immunoassay. The enzyme-linked immunosorbent assay method utilizes a synthetic paraoxon-methyl immunogen for immunization, thereby obtaining a polyclonal antibody; the paraoxon-methyl is taken as a standard product, and a conjugate of the paraoxon-methyl hapten and OVA is taken as a coating antigen, thereby establishing the indirect competitive enzyme-linked immunosorbent assay method of the paraoxon-methyl in textiles (cotton). The enzyme-linked immunosorbent assay method establishes the indirect ELISA method of the paraoxon-methyl pesticide metabolite in the textiles, thereby providing the rapid and high-efficient detection means for the residual detection of the paraoxon-methyl in the textiles; as the enzyme-linked immunosorbent assay method adopts the polyclonal antibody, the cost is lower, the stability and the repeatability are better. The sensitivity is 0.01ppm and the linear range is 0.1 to 20ppm. The high specificity and the high affinity of the immune reaction allow the ELISA to have very high selectivity and sensitivity.

The invention relates to a rhodamine B ELISA detection method, which belongs to the technical field of ELISA. The invention discloses a rhodamine B ELISA detection method, which comprises the following steps: using synthesized rhodamine B immunogen for immunizing healthy New Zealand rabbits to obtain polyclonal antibodies; and using rhodamine B as standard products; using rhodamine B hapten and OVA coupling materials as coating antigen to establish a rhodamine B indirect ELISA method. The invention provides the fast and efficient detection method for the residual detection on the rhodamine B. Because the polyclonal antibodies are adopted, the cost is low, and in addition, the stability and the repetitiveness are good. The detectability (IC90) is 0.028 ng/mL, the half inhibitory amount (IC50) is 1.3 ng/mL, and the detection range (IC20 to IC80) is between 0.07 and 17.5 ng/mL. Because of high specificity and high compatibility of the immune reaction, high selectivity and high sensitivity can be obtained when the ELISA is used for detecting the rhodamine B.

The invention discloses a PEDV S gene major antigen epitope serial connection recombination gene, and a preparation method and an application thereof, and belongs to the field of a PEDV S protein major neutralizing epitope region efficient expression method. The PEDV S gene major antigen epitope serial connection recombination gene is obtained through connecting three major antigen epitopes of a PEDV S gene in series; every two major antigen epitopes are connected through a flexible amino acid base sequence; the name is S123; and the sequence of the base sequence is shown as SEQ ID NO:1. The recombination gene contains most PEDV antigen epitopes; the selected fragments avoid a high-variation region; the expression quantity is efficient and stable; the immunogenicity is high; when the application of PEDV S gene major antigen epitope serial connection recombination protein to a PEDV indirect ELISA detection kit is used, the PEDV can be fast diagnosed; and the swinery neutralization protection can be well monitored and evaluated.

The invention relates to virus-like particle recombinant protein of a virus variation strain VP2 gene of an infectious bursal disease, belonging to the field of biologic pharmacy. The IBDV variation strain (AH1) VP2 gene is cloned, converted and transfected to obtain a recombinant baculovirus vBac-VP2; an infected Sf9 insect cell has specific fluorescence, and the antigen valence of the infected Sf9 insect cell is above 1.6*10<3>; the molecular weight of a recombinant VP2 protein is 53kDa, and the recombinant VP2 protein is in the state of virus-like particles; an indirect ELISA detection method established for an envelope antigen by the purified recombinant VP2 protein has good specificity and sensibility; an immune chicken can resist IBDV virulent attack, and the protection ratio achieves 100 percent. The novel virus-like particle recombinant VP2 protein prepared by the IBDV variation strain VP2 gene has high pertinence on the immune prevention of a current prevalent IBDV virulent strain and good practical value and popularization prospects.

This invention discloses monoclonal antibody against anti-LCDV immunoglobulin of Paralichthys olivaceus, which is excreted by hybridoma JF-lgM-H (CCTCC-C200631). The method comprises: immunizing Paralichthys olivaceus with LCDV inactivated by formalin to prepare antiserum, purifying Paralichthys olivaceus immunoglobulin, immunizing Balb/c mice as antigen, preparing hybridoma cells by cell engineering method, and screening the monoclonal antibody by immunoassay. Indirect ELISA and indirect immunofluorescent antibody assay show that this monoclonal antibody is located on the heavy chain (7-80 kDa) of the anti-LCDV immunoglobulin. The monoclonal antibody can be used for preparing reagents for detecting LCDV infection in early stage, and evaluating the immune effects of LCDV vaccine inactivated by formalin.

The invention discloses an indirect ELISA kit for detecting African swine fever virus (ASFV) antibody, and a use thereof, and belongs to the technical field of biology. The kit is characterized by: adopting prokaryotic expressed recombined P54 protein as coating antigen; detecting antibody against to the ASFV in swine serum according to an indirect ELISA principle. The coating antigen in 96-well plates of the kit is the prokaryotic expressed recombined P54 protein which has good antigenicity. The enzyme-linked immunospecific assay kit provided by the present invention comprises the 96-well plates coated by the P54 protein, positive control, negative control, horseradish peroxidase-labeled rabbit anti-porcine IgG polyclonal antibody, a concentrated cleaning solution, a serum dilution, a TMB substrate indicator and a stop buffer. The kit provided by the present invention can be provided for screening mass samples. In addition, the main reagents in the kit are provided in a working fluid manner such that the reagents are used conveniently.

The invention discloses a universal phthalic acid esters monoclonal antibody hybridoma cell strain and application thereof, and belongs to the technical field of food safety immunological detection. Hapten is prepared and is coupled with albumen based on the glutaraldehyde method to obtain phthalic acid dibutyl ester complete antigen, and the phthalic acid dibutyl ester complete antigen and freund's adjuvant are evenly mixed to be injected to immune BALB/c mice in a subcutaneous injection mode; envelope antigen is formed in a synthesis mode based on the diazotization method and is used for screening mouse serum and cell supernatant. The splenocyte of the immune mice is fused with myeloma cells of the mice based on a PEG method, and indirect ELISA, indirect competition ELISA screening and three times of subcloning are carried out to obtain selective group hybridoma cell strain monoclonal cell strain C. The monoclonal cell strain C has certain recognition capability on DEHP and DINP, and the requirement for phthalic acid ester plasticizer immunodetection products in current market can be met.