151 results about "Coating antigen" patented technology

The invention provides an ELISA (Enzyme Linked Immunosorbent Assay) kit for detecting an anti-MDA5 antibody and a preparation method of the ELISA kit. According to the kit, full-length human MDA5 protein (1025 amino acids) expressed by a cell-free protein expression system under the low-temperature condition of 22 DEG C is adopted as a coating antigen, and a disulfide bond enhancer E6820 is added into the system to promote correct folding of the protein. The kit comprises a microwell plate coated with a full-length MDA5 antigen, an enzyme-labeled anti-human IgG secondary antibody, a standard substance with the concentration range of 1-243 ng / mL, a chromogenic substrate and an optimized confining liquid system. The invention solves the problem of insufficient sensitivity caused by the use of fragment antigens in the prior art, and can be used for early diagnosis and illness monitoring of dermatomyositis-related interstitial lung diseases.

The invention provides a fusion protein for detecting an aquaporin 4 antibody as well as a preparation method and application of the fusion protein, and belongs to the technical field of aquaporin 4 antibody detection. The nucleotide sequence of the fusion protein is shown as SEQ ID NO.5. The fusion protein is used as a coating antigen, M1 type and M23 type aquaporin 4 can be detected at the same time, and the sensitivity and the accuracy of detecting an aquaporin 4 antibody are high, and the repeatability is good. When the kit is used for the aquaporin 4 antibody, the detection method is simple and rapid, and the kit is suitable for clinical and scientific research large-scale screening of the aquaporin 4 antibody.

The invention discloses an indirect ELISA (Enzyme-Linked Immunosorbent Assay) detection kit for distinguishing host infected Listeria monocytogenes and / or Listeria ivanovii. The coating antigens are respectively a recombinant listeria monocytogenes surface protein ActA, a serotype 4h listeria monocytogenes and listeria ivanovii common outer membrane protein SmcL and a listeria ivanovii specific outer membrane protein I-inlH, indirect ELISA methods are respectively established based on the three antigens, the three detection methods are combined to detect the content of the listeria monocytogenes, and the content of the listeria ivanovii can be detected. The levels of the ActA antibody, the SmcL antibody and the I-inlH antibody in a clinical serum sample can be effectively monitored, and the detection on the listeria monocytogenes caused by the listeria monocytogenes and / or the listeria ivanovii is realized. The kit is relatively high in sensitivity, strong in specificity and good in stability, and an effective method is provided for epidemiological investigation and diagnosis of listeria monocytogenes.

The present invention belongs to the field of biotechnology, and relates to a hybridoma cell strain secreting a monoclonal antibody of Clostridium perfringens CPA with neutralizing activity and its application. The present invention uses natural CPA as an immune antigen and a non-toxic CPA mutant as a coating antigen for the first time, and screens a hybridoma cell strain secreting a monoclonal antibody with neutralizing activity. The hybridoma cell strain can stably and efficiently secrete a monoclonal antibody with neutralizing activity, and can achieve large-scale batch production, thereby providing a material basis for the detection and treatment of CPA antigen / antibody in the later stage.

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 invention provides an infectious bronchitis virus antigen epitope fusion peptide, and the prepared antigen epitope fusion peptide can be used for preparing an antibody and can also be used as a coating antigen for preparing an indirect ELISA detection product. The amino acid sequence of the provided epitope fusion peptide is SEQ ID NO: 4. After antigen epitope analysis is performed on S1 protein of an infectious bronchitis virus strain through bioinformatics software, a plurality of antigen epitopes are fused to obtain the antigen epitope fusion peptide. Specificity tests show that the prepared antigen epitope fusion peptide is a coating antigen, does not have cross reaction with positive serum of other common pathogens, and only has good reaction with positive serum of GI-19 type IBV, that is, the antigen epitope fusion peptide has good serotype specificity as a detection antigen, and has small cross reaction with non-GI-19 type IBV antibody.

The invention discloses a liquid-phase protein chip method for simultaneously detecting three kinds of porcine coronaviruses and a kit thereof, which comprises the following steps of: after activating and coupling microspheres, respectively coating antigen on porcine delta coronavirus N protein, porcine acute diarrhea syndrome coronavirus N protein and porcine epidemic diarrhea virus N protein by using the microspheres; matching the two fluorescent dyes according to different concentrations to form fluorescent coding microsphere combination arrays with different energy levels; the microspheres are analyzed one by one based on the flow principle, red laser recognizes microsphere codes, green laser reads fluorescence intensity of report molecules, computer analysis and processing are carried out, and fluorescence signals are converted into quantitative detection results; and determining the optimization of the detection conditions of the liquid-phase protein chip after the liquid-phase protein chip test. The antigens coated on the microspheres only specifically react with corresponding antibodies and are not combined with antibodies generated by other pathogens; the PDCoV, SADS-CoV and PEDV antibodies in the sample to be detected can be rapidly, effectively and accurately detected, and reference is provided for prevention of the porcine coronavirus.

The invention relates to the technical field of biochemical detection, in particular to an enzyme linked immunosorbent assay kit for detecting diisononyl phthalate and application of the enzyme linked immunosorbent assay kit. The invention relates to an enzyme linked immunosorbent assay kit for detecting diisononyl phthalate. The enzyme linked immunosorbent assay kit comprises the following components: an elisa plate coated with a coating antigen, a diisononyl phthalate specific antibody working solution, an enzyme marker working solution, a diisononyl phthalate standard substance solution, a substrate developing solution, a stopping solution, a washing solution, a sample diluting solution and a purifying agent. According to the invention, the content of diisononyl phthalate in an edible oil sample is qualitatively or quantitatively detected by mainly adopting an indirect competitive ELISA method; the sample pretreatment requirement is low, the sample pretreatment process is simple, and a large batch of edible oil samples can be rapidly detected at the same time; the main reagent is provided in the form of a working solution, and the detection method is convenient and easy to implement and has the characteristics of high specificity, high sensitivity, high precision, high accuracy and the like.

The invention relates to the technical field of biological products, in particular to an indirect ELISA (Enzyme-Linked Immunosorbent Assay) detection kit and a detection method for a goose astrovirus antibody. The kit comprises an elisa plate, an elisa secondary antibody, goose astrovirus positive control serum, goose astrovirus negative control serum, a diluent, a washing solution, a developing solution and a stop solution, wherein a coating antigen of the ELISA plate is a goose astrovirus VP27 recombinant protein, the goose astrovirus is a goose astrovirus GAstV / SD / 36, and the goose astrovirus VP27 recombinant protein corresponds to an ORF2 region. The goose astrovirus GAstV / SD / 36 is used as a template, the virus antigen recognition site VP27 protein is selected as an antigen for preparing the indirect ELISA kit, the indirect ELISA detection kit of the goose astrovirus antibody is prepared, and the accuracy of the detection result of the kit can be improved.

The invention relates to the field of anti-autologous AchR antibody detection equipment, in particular to a method for efficiently detecting an anti-autologous AchR antibody, which comprises the following operation steps: S1, antigen coating: selecting a proper microwell plate, diluting an AchR antigen to a proper concentration by using a carbonate buffer solution, adding 100-200 [mu] L of an antigen solution into each well, then placing the microwell plate in a refrigerator at 4 DEG C overnight, and taking the microwell plate out of the refrigerator; the antigen can be fully adsorbed on the surface of the microwell plate; s2, closing the microwell plate: after overnight coating, pouring away the antigen solution in the microwell plate, washing the microwell plate with a washing solution for three times, soaking for 3-5 minutes each time to remove uncombined antigens, then adding 200-300 microliters of a sealing solution into each hole, incubating for 1-2 hours at 37 DEG C, and after incubation is finished; according to the invention, the anti-autologous AchR antibody is detected based on the highly specific binding reaction of the antigen and the antibody, and the layer-by-layer specific binding mechanism enables the ELISA to have high specificity on the detection of the target antibody, so that the interference caused by non-specific binding can be effectively reduced.

The invention belongs to the technical field of detection, and particularly relates to a universal kit for bovine nodular skin viruses, sheep pox viruses and goat pox viruses. On the basis of expressed specific recombinant p122 protein and serum to be detected as a detection sample, the double-antigen sandwich ELISA method for the bovine nodular skin virus, the sheep pox virus and the goat pox virus is established, and the method comprises the following steps: coating an antigen, sealing, adding the sample to be detected and an enzyme-labeled antigen, developing, judging a result and the like. The kit has the advantages of being good in specificity, high in sensitivity, good in repeatability, low in cost and capable of achieving batch detection, and a certain detection technology foundation can be laid for preventing and controlling the bovine nodular skin virus, the sheep pox virus and the goat pox virus.

The invention discloses a recombinant CapA protein of cat astrovirus. The recombinant CapA protein has an amino acid sequence as shown in SEQ ID NO.1 in a sequence table or is coded by a base sequence as shown in SEQ ID NO.2 in the sequence table. Therefore, a cat astrovirus indirect ELISA antibody detection kit is developed and comprises a coated ELISA plate, and the coated ELISA plate takes the recombinant CapA protein as a coating antigen. Meanwhile, a corresponding indirect ELISA antibody detection method is established. Research results show that the critical value of the indirect ELISA antibody detection method and kit is 0.523; the sensitivity is 1: 32000; the in-batch variation coefficient of detected samples is 2.679%-7.910%, the inter-batch variation coefficient is 3.627%-8.982%, both the in-batch variation coefficient and the inter-batch variation coefficient are smaller than 10%, and the clinical application effect is good. Therefore, the kit has the characteristics of high sensitivity, strong specificity, good repeatability and the like, can quickly and effectively detect the FAstV antibody, and provides technical support for accurate diagnosis, timely prevention and control and vaccine effect evaluation of FAstV.

The invention discloses an indirect ELISA (Enzyme-Linked Immunosorbent Assay) kit for rapidly detecting a porcine delta coronavirus IgA antibody, and relates to the field of immunological detection. The indirect ELISA kit comprises an ELISA plate coated with an antigen, the antigen is a recombinant PDCoV S2 protein, and the amino acid sequence of the antigen is as shown in SEQ ID NO. 2. According to the indirect ELISA kit, a recombinant PDCoV S2 protein is used as a coating antigen to capture a PDCoV specific IgA antibody, a goat anti-pig IgA antibody labeled by horse radish peroxidase is used as a detection antibody, a 'PDCoV-S2 protein-PDCoV-S2 specific IgA antibody-enzyme labeled antibody' trimer compound can be formed on an ELISA plate, and the PDCoV specific IgA antibody-enzyme labeled antibody can be used for detecting the PDCoV specific IgA antibody. And the result can be directly judged by measuring the color change of the substrate and the absorbance value, so that the detection efficiency can be obviously improved. The indirect ELISA kit can be used for clinical detection and immune evaluation of the PDCoV.

The invention discloses a method for establishing duck hepatitis virus antibody ELISA (enzyme-linked immuno sorbent assay) detection, which comprises the following steps: firstly, preparing experimental materials, and then cloning and antigenicity analysis of structural genes, inducible expression of positive recombinants in escherichia coli, expression and purification of recombinant proteins, antigenicity analysis of the recombinant proteins and establishment of a recombinant antigen VP3 indirect ELISA method. Finally, VP0, VP3 and VP1 recombinant proteins can react with duck anti-DHV-1 positive serum and have good antigenicity, indirect ELISA results show that under the same coating concentration, the P / N value of the fusion protein GST-VP3 serving as a coating antigen is higher than that of the fusion protein GST-VP3 serving as the coating antigen and that of the fusion protein GST-VP3 serving as the coating antigen, it is indicated that the antigenicity of the fusion protein GST-VP3 is better than that of the fusion protein GST-VP3 serving as the coating antigen and that of the fusion protein GST-VP3 serving as the coating antigen and that of the fusion protein GST-VP3 serving as the coating antigen and that of the fusion protein GST-VP3 serving as the coating antigen and that of the fusion protein and the result is consistent with the prediction result.

The invention discloses a detection method and a kit for detecting listeria monocytogenes, and relates to the technical field of biology. The kit is an indirect ELISA kit and comprises an elisa plate coated with an antigen, the amino acid sequence of the antigen is as shown in SEQ ID NO. 2. The indirect ELISA detection kit constructed by the invention only reacts with listeria monocytogenes positive serum, has no cross reaction with other serum, and has strong specificity; when the serum dilution is 1: 800, the detection result is still positive, and the sensitivity is high; the intra-batch variation coefficient ranges from 2.79% to 6.64%, the inter-batch variation coefficient ranges from 2.61% to 7.75%, and the intra-batch repeatability and the inter-batch repeatability are good; 24 serum samples are detected, and the total coincidence rate is 95.83%. The kit provided by the invention can effectively detect sample infection of Listeria monocytogenes, and has the advantages of strong specificity, high sensitivity and good repeatability.

The application relates to a sensor for synchronously detecting enniatin and roquefortine based on a double-potential electrochemiluminescence immunoassay technology as well as a preparation method and a detection method thereof. The sensor comprises a base electrode, a sensing interface composed of a nanocomposite is modified on the surface of the base electrode, mixed coated antigens including ENNs-BSA and AOH-BSA fixed on the sensing interface, mixed probes including a cathode probe and an anode probe, the cathode probe is Zr-Por-MOF-ENNs Ab which is obtained by marking ENNs antibodies with a porphyrin zirconium metal organic framework, the anode probe is Ce-In-MOF@ABEI-AOH Ab which is obtained by marking AOH antibodies with ABEI loaded on a cerium-doped indium metal organic framework, and a detection solution containing a single co-reactant potassium persulfate. The application can realize one-time sample adding, double-potential scanning and synchronous quantification for two kinds of mycotoxins of enniatin and roquefortine.

This invention discloses a quantitative test strip for HBV PreS1 antibody based on the AIE molecule, its preparation method, and its applications, relating to the fields of molecular biology, immunology, and biodetection technology. The test strip includes a sample pad, a labeling pad, a nitrocellulose membrane (NC membrane), an absorbent pad, and a base plate. The NC membrane detection line is coated with HBV PreS1 antigen to capture PreS1 antibodies in the sample. The labeling pad contains an immobilized Anti-Human IgG antibody conjugated to the AIE molecule, which binds to the PreS1 antibodies in the sample to form an immune complex. The test strip of this invention enables rapid detection of PreS1 antibodies and achieves enhanced fluorescence in the detection results. Combined with a fluorescence reader or fluorescence card reader, the detection results can be digitally and quantitatively analyzed. It is simple to operate, rapid in detection, and significantly improves detection sensitivity and enables quantitative analysis.

The application relates to a detection method of anti-preS1 antibody, an anti-preS1 antibody detection kit and application, the application carries out multi-sequence comparison and conservation analysis on preS1 sequences of different genotypes (A, B, C, D and CD mixed type) of HBV, takes type A (AB697487.1), type C (FJ032351.1) and type D (LC519808.1) as reference sequences, based on the sequence consistency standard of 85% of full length and 90% of the key region of functional antibody binding, a highly conservative preS1 full-length antigen sequence is systematically screened out in natural variation, the sequence can cover antigen variation of A-D type HBV, and the sequence is taken as a coating antigen to establish an indirect ELISA detection system, a good linear relationship is presented in the range of 4.88-625 ng / mL, and the Cut-off value is 33.6 ng / mL; clinical verification shows that the method can effectively distinguish chronic hepatitis B patients from healthy control groups, compared with detection methods based on partial fragments or overlapping short fragments, the false negative rate can be significantly reduced, and the method is suitable for HBV infection immune state evaluation, anti-virus curative effect monitoring and prognosis analysis.

The present invention relates to the field of diagnostic detection of animal virus antibodies, and specifically provides a method for detecting African swine fever virus antibodies and its application. First, the present invention screens the dominant epitope regions of the structural proteins p30, p54, and p72 of African swine fever, tandemly expresses and purifies the three dominant epitope regions using an Escherichia coli expression system, and uses the purified protein as a coating antigen to establish an indirect ELISA method for detecting clinical swine sera. The present invention makes up for the deficiencies of existing domestic and foreign detection methods, and will be more conducive to the monitoring of antibody levels and prevalence generated by ASFV vaccine immunization and wild virus infection.

The invention relates to an indirect ELISA (enzyme-linked immuno sorbent assay) detection kit for a sheep bacteroides nodules antibody and application thereof, belonging to the technical field of biological detection. In order to solve the technical problems that a bacterium separation identification and PCR (Polymerase Chain Reaction) detection method of bacteroides nodulosus has defects and a serological identification method for bacteroides nodulosus does not exist in the prior art, the invention adopts a sheep bacteroides nodulosus outer membrane protein Omp1D as a coating antigen in an indirect ELISA (Enzyme-Linked Immuno Sorbent Assay) detection kit; rapid detection and screening of bovine and sheep bacteroides nodules infection can be realized, and the kit has the characteristics of simple operation, rapidness and sensitivity, accurate result, good specificity and repeatability and the like; meanwhile, the indirect ELISA detection kit for the sheep bacteroides nodules antibody has the characteristics of being simple in preparation method, low in cost, suitable for large-scale production and the like, a rapid, simple and sensitive diagnostic kit for detecting bacteroides nodules infection is provided for large-scale breeding of animal husbandry, and a technical support is provided for preventing and controlling bacteroides nodules infection.

This invention discloses an indirect ELISA detection kit and method for bovine pasteurellosis, relating to the field of biotechnology. The indirect ELISA kit includes an enzyme-labeled plate coated with an antigen; the amino acid sequence of the antigen is shown in SEQ ID NO.2. The indirect ELISA detection kit constructed in this invention exhibits high specificity and sensitivity; the intra-assay coefficient of variation is between 1.79% and 7.54%, and the inter-assay coefficient of variation is between 2.23% and 6.91%, both below 10%, demonstrating good inter-assay and intra-assay repeatability. In testing 30 clinical samples, the overall concordance rate with the standard "Diagnostic Techniques for Bovine Hemorrhagic Sepsis (GB / T27530-2025)" was 96.67%. The indirect ELISA detection kit demonstrates good overall performance and can be applied to the detection of bovine pasteurellosis in clinical serum samples.

The invention provides a linear epitope of an African swine fever virus CP312R protein. The amino acid sequence of the linear epitope is SEQ ID NO: 1 or SEQ ID NO: 2. The antigen epitope polypeptide provided by the invention has good antigen specificity and immunoreactivity. The polypeptide can be efficiently combined with an African swine fever virus specific antibody, background interference is low, cross reaction is small, and detection sensitivity and accuracy are remarkably improved. By using the polypeptide as a coating antigen in immunodetection methods such as ELISA or colloidal gold immunochromatography, rapid and accurate detection of ASFV infection can be realized, and the detection result is stable and reliable. Compared with a traditional whole virus antigen or a crude extract antigen, the epitope polypeptide has the advantages of being simple and convenient to prepare, small in batch difference, high in safety and the like, and is suitable for ASFV serological monitoring and diagnosis application.

The invention discloses a method and a kit for simultaneously detecting avian leukosis virus, mycoplasma synoviae and salmonella pullorum antibodies, and belongs to the technical field of animal epidemic disease detection. According to the method, an avian leukosis virus p27 protein, a chicken mycoplasma synoviae p50 protein and a truncated GroEL-delta8-1 protein of a dominant antigen GroEL of salmonella pullorum are taken as coating antigens, synchronous detection of the three antibodies is realized by optimizing ELSA reaction conditions, and a kit capable of simultaneously detecting the three antibodies is developed at the same time. The kit has the characteristics of strong specificity, high sensitivity and simplicity and convenience in operation, is suitable for rapid screening and purification of poultry epidemic diseases, and reduces the detection cost.

The invention discloses a polypeptide-based avian reticuloendotheliosis virus indirect ELISA antibody detection method, and belongs to the technical field of biology. According to the kit, potential epitopes of REV gp90 protein are systematically screened to obtain three sections of specific polypeptide antigens for detecting the avian reticuloendotheliosis virus antibody, and a mixed product of the three sections of specific polypeptide antigens is used as a coating antigen to develop the indirect ELISA antibody detection kit for the avian reticuloendotheliosis virus. On the basis of the indirect ELISA antibody detection kit, the avian reticuloendotheliosis virus antibody detection method established by optimizing the coating concentration, the sealing condition, the serum incubation time and the like has the advantages of strong specificity, high sensitivity, good repeatability and convenience in operation; the kit can effectively and rapidly diagnose epidemic diseases caused by the avian reticuloendotheliosis virus, and is beneficial to popularization and application.

This invention discloses a kit for detecting porcine reproductive and respiratory syndrome virus (PRRSV) antibodies and its applications, belonging to the field of animal disease detection technology. The kit uses a mutant nucleocapsid protein (N protein) of the PRRSV-2 class NADC30 strain CHsx1401 as the coating antigen; the N protein mutant has mutations at amino acid positions 23, 75, and / or 90 compared to the wild-type N protein. High purity is achieved through optimized prokaryotic expression systems and purification processes. Under optimal detection conditions, the kit exhibits strong specificity, high sensitivity, and good reproducibility, with significantly superior detection performance compared to existing commercial kits. It can be used for PRRSV-2 antibody detection, epidemiological surveys, and disease control.

This invention constructs pET-32a-CP1 and pET-32a-CP2 expression vectors based on the CP1 and CP2 protein genes of Giardia duodenalis, transforms them into Rosetta (DE3) competent cells, and induces expression using IPTG. After SDS-PAGE analysis of the recombinant protein expression, Ni-NTA affinity chromatography is used to purify the proteins. Two indirect ELISA methods are then established using these as coating antigens. The indirect ELISA methods based on the recombinant proteins CP1 and CP2 of this invention exhibit good specificity, sensitivity, and repeatability, with concordance rates of 98% and 94% with microscopic detection results, respectively.

The invention discloses an OVA-D fluorescence immunochromatography detection test strip in a surimi product and application thereof, the test strip comprises a substrate, a sample adding pad, a combination pad, a reaction film and a water absorption pad, the sample adding pad, the combination pad, the reaction film and the water absorption pad are arranged on the substrate, the reaction film is arranged in the middle of the substrate, one end of the sample adding pad is in contact with the other end of the combination pad, one end of the combination pad is in contact with the other end of the reaction film, and the other end of the reaction film is in contact with the water absorption pad. One end of the reaction film contacts with the other end of the water absorption pad; the combination pad contains a rabbit anti-OVA-D polyclonal antibody marked by fluorescent quantum dots and goat IgG marked by fluorescent quantum dots; the reaction film is provided with a detection area, the detection area is sequentially provided with a detection line and a control line from the other end to one end of the reaction film, the detection line is coated with an OVA-D antigen, and the control line is coated with a rabbit anti-goat IgG antibody. The fluorescence signal is stable, the fluorescence intensity of the T line and the C line is not influenced by a complex matrix in a detected sample, the detection sensitivity is higher, and the reliability of quantitative detection of the duck egg ovalbumin is also ensured.

The invention discloses a method for detecting malachite green, crystal violet and methylene blue, which is characterized in that two different haptens are respectively used as an immunogen and a coating antigen to obtain a high-sensitivity monoclonal antibody for resisting malachite green, crystal violet and methylene blue. The invention further establishes a chemiluminescence immune method for detecting malachite green, crystal violet and methylene blue based on biotin-streptavidin, and the method has the advantages of rapidness, sensitivity, low cost and high throughput, and is suitable for simultaneously detecting the residual quantity of malachite green, crystal violet or methylene blue in aquatic products.

The invention belongs to the technical field of biology, and particularly relates to a bovine viral diarrhea virus NS2 truncated protein and application thereof in BVDV antibody detection. The invention firstly provides a truncated protein of a BVDV NS2 protein, the truncated protein is subjected to soluble expression after being fused with glutathione S transferase (GST) and horse radish peroxidase (HRP) respectively, has better reactogenicity with positive serum of a BVDV antibody, does not react with negative serum of the BVDV antibody, and can be used for detecting the BVDV antibody; secondly, a fusion protein of GST and the truncated protein is taken as a coating antigen, a fusion protein of HRP and the truncated protein is taken as a detection antigen, a double-antigen sandwich ELISA method for BVDV antibody detection is constructed, and the method has the technical advantages of high sensitivity, good specificity, simplicity and convenience in operation, easiness in popularization, low cost and the like, and can be applied to screening of BVDV antibodies in cattle.

The invention provides an indirect ELISA (Enzyme-Linked Immunosorbent Assay) antibody detection method and an indirect ELISA antibody detection kit based on pyruvate Kinase (PK), and relates to the technical field of parasite detection. The kit comprises a coating antigen Italiella mannici PK protein, a primary antibody, a labeled secondary antibody and an initiator solution, wherein the primary antibody is a mouse or cat anti-Italiella mannici PK protein antibody to be detected; the labeled secondary antibody can be combined with the primary antibody combined with the PK protein of the andrometrium mansoni; and the labeled second antibody is provided with a detection marker. The method comprises the following steps: cloning a PK protein gene into a prokaryotic expression vector PQE80L (+), constructing a recombinant plasmid PQE80L-PK, inducing PK recombinant protein expression by utilizing IPTG (isopropyl-beta-d-thiogalactoside), purifying the recombinant protein to coat an elisa plate, and establishing an indirect ELISA (enzyme-linked immuno sorbent assay) detection method of the Mangnocorynth PK protein antibody by optimizing reaction conditions. The detection method has the advantages of high sensitivity and strong specificity, and provides support for the development of the detection technology of the sitonia mansoni and the prevention and control of the schizocyst.