13 results about "Indirect elisa" patented technology

The application belongs to the technical field of biology and particularly relates to an African swine fever virus pNP868R protein monoclonal antibody and a preparation method and application thereof. The application takes the pNP868R protein prepared by eukaryotic expression as an antigen, immunizes BALB / c mice, uses hybridoma technology, and obtains a hybridoma cell line capable of stably secreting the anti-African swine fever virus pNP868R protein monoclonal antibody through cell fusion, indirect ELISA method screening, limited dilution method subcloning and hybridoma continuous passage culture. The application also obtains an African swine fever virus pNP868R protein monoclonal antibody. The monoclonal antibody has high affinity with the pNP868R protein and can be applied in Western-Blot experiments, indirect immunofluorescence experiments, immunoprecipitation experiments and the capture of proteins interacting with pNP868R, and can be used for the diagnosis, prevention or treatment of African swine fever.

This invention relates to the application of the avian adenovirus serotype 4 multi-epitope recombinant chimeric protein rcF2 in the preparation of a kit for detecting avian adenovirus serotype 4 antibodies. The amino acid sequence of the avian adenovirus serotype 4 multi-epitope recombinant chimeric protein rcF2 is shown in SEQ ID NO.1. This invention utilizes the multi-epitope recombinant chimeric protein rcF2 to establish an indirect ELISA detection method and kit for detecting avian adenovirus serotype 4. This recombinant chimeric protein can be recognized by avian adenovirus serotype 4 positive sera, and the detection sensitivity of positive sera samples can reach 1:16000. It shows no cross-reactivity with antibodies against other pathogens, indicating that the ELISA detection method based on this multi-epitope recombinant chimeric protein rcF2 is simple to operate, enhances detection sensitivity, and has good specificity and repeatability.

The application belongs to the field of biology, discloses PDCoV recombinant RBD protein and IgA ELISA kit thereof, and the amino acid sequence of the recombinant RBD protein is shown as SEQ ID NO.1.The recombinant RBD protein is used for indirect ELISA kit, can cover early IgA response of infection, can accurately recognize PDCoV specific IgA antibody in breast milk sample, and will not cause false positive due to infection of other common pathogens in a pig herd; the indirect ELISA kit of the application is stable (the batch variation coefficient is 1.848%~6.062%, and the batch variation coefficient is 4.504%~7.790%), and the coincidence rate with an immunofluorescence experiment reaches 93.26%, the repeatability and the coincidence rate are high, the method has the advantages of simple operation, short time consumption and large detection flux, and is suitable for large-scale screening of clinical pig milk samples.

The present invention belongs to the technical field of biological detection, and in particular to an indirect ELISA detection kit for infectious bovine rhinotracheitis virus (IBRV) antibodies and use thereof. Based on the good antigenicity of an IBRV gE protein, the present invention endows the indirect ELISA detection kit and an indirect ELISA detection method for IBRV, established based on the IBRV gE protein, with characteristics of sensitivity, specificity, and high efficiency. The method is simple to operate, time-efficient, and cost-effective, allowing for high-throughput detection of cattle infected with IBRV, and provides a simple and effective diagnostic tool for clinical diagnosis of infectious bovine rhinotracheitis (IBR).

The application discloses a monoclonal antibody of a porcine rotavirus VP6 protein, an epitope of the monoclonal antibody and application of the monoclonal antibody. The monoclonal antibody can specifically recognize a linear B cell epitope on the VP6 protein, and a core sequence of the epitope is TTRLSFQLVR. The antibody has high affinity and high specificity, and can cross-react with porcine rotaviruses of various genotypes such as G5, G9 and G11. The application further includes an indirect ELISA detection kit containing the monoclonal antibody, and application of the linear epitope in detection of porcine rotaviruses, development of vaccines and research on functions of viral proteins. The application provides an important tool and theoretical basis for serological diagnosis, vaccine evaluation and etiological research of porcine rotaviruses.

The application discloses a Mycobacterium avium subsp. paratuberculosis (MAP) indirect ELISA antibody detection kit and application thereof, and belongs to the technical field of microorganism detection. The kit contains a microplate coated with a recombinant protein MAP 625472 composed of three proteins MAP1272c, MAP0862 and MAP2154c in series. Experiments prove that the indirect ELISA method based on the recombinant protein MAP 625472 has high accuracy, high sensitivity and high specificity. In the detection of clinical serum, the positive coincidence rate of the kit is 96.7%, the negative coincidence rate is 95.1%, and the sensitivity is higher than that of an IDvet paratuberculosis antibody detection kit, indicating that the MAP indirect ELISA antibody detection kit can replace foreign products for the detection of bovine PTB, and provides an effective technical means for the prevention and control of bovine PTB in China.

The application discloses a porcine eperythrozoon recombinant MSG1 protein and an indirect ELISA antibody detection kit prepared from the porcine eperythrozoon recombinant MSG1 protein, and a preparation method and application of the kit, wherein the kit comprises a support medium coated with the porcine eperythrozoon recombinant MSG1 protein, an enzyme label, and a detection reagent for porcine eperythrozoon antigen antibody, an antibody diluent, a negative control and a positive control. The application realizes efficient soluble expression of the porcine eperythrozoon MSG1 antigen, avoids the complexity of inclusion body renaturation, reduces the cost and retains the natural immunogenicity of the protein. The established ELISA detection method is systematically optimized, has high sensitivity and specificity, is suitable for large-scale serological screening and epidemiological monitoring, and provides a reliable tool for primary prevention and control.

The application discloses a polypeptide for detecting type 1 RHDV antibody, and an amino acid sequence of the polypeptide is shown as SEQ ID NO. 1. The polypeptide is applied to preparing a kit for detecting type 1 RHDV antibody and is applied to detecting type 1 RHDV antibody. The application further discloses a kit for detecting type 1 RHDV antibody, which comprises the polypeptide for detecting type 1 RHDV antibody. The kit for detecting type 1 RHDV antibody is applied to detecting type 1 RHDV antibody. The polypeptide for detecting type 1 RHDV antibody has high specificity for type 1 RHDV antibody, is safe and has good antigenicity. The kit for detecting type 1 RHDV antibody is suitable for an indirect ELISA detection method, can evaluate type 1 RHDV antibody level, has high specificity and sensitivity and is suitable for detecting batch samples.

The application discloses a pig SLPI monoclonal antibody and application thereof, and belongs to the field of biotechnology and cell engineering. The application immunizes mice with a His-SLPI recombinant protein, and carries out cell fusion when the serum titer reaches 1:10000 or above. On the seventh day of the fusion, indirect ELISA is carried out on cell supernatant, and positive wells with higher OD values are screened out. The purified recombinant GST-SLPI protein is used as a coating antigen, the wells are expanded and cultured, and the cells are made into single cells in single wells by using an effective dilution method. After continuous screening to obtain 100% positive wells, the supernatant of the positive cells is used for specific identification of the pig-derived SLPI protein by Western blot and IFA. One hybridoma strain B with a higher binding titer is obtained by identification, the heavy chain amino acid sequence of the SLPI monoclonal antibody produced by the hybridoma strain B is shown as SEQ ID NO. 1, and the light chain amino acid sequence is shown as SEQ ID NO. 2.

This invention relates to the field of biomedicine, specifically to an antibody for detecting Escherichia coli O45 and its application method. The antibody is a monoclonal antibody or its antigen-binding fragment, comprising the heavy chain CDR sequence shown in SEQ ID NO:1-3 and the light chain CDR sequence shown in SEQ ID NO:4-6. This invention also provides the nucleic acid molecule encoding the antibody, the expression vector, the host cell, and the preparation method thereof. Specificity identification results show that the antibody specifically binds to Escherichia coli O45 and shows no cross-reactivity with other Escherichia coli serotypes such as O157, O26, O103, O111, O121, and O145, as well as closely related serotypes of O55, and also shows no cross-reactivity with common foodborne pathogens. The indirect ELISA detection method based on this antibody has a detection limit of 1×10⁻⁶ for Escherichia coli O45. 2 The detection rate is CFU / mL, and the entire testing process can be completed within 24 hours. The monoclonal antibody provided by this invention can be used to prepare detection kits, test strips, and other products, and is suitable for rapid screening of O45 Escherichia coli in imported and exported food.

This invention provides a method for preparing a bovine coronavirus monoclonal antibody and its corresponding antigenic epitope peptide. The monoclonal antibody is prepared by immunizing animals with the N protein of bovine coronavirus as an antigen, followed by screening hybridoma cells. The amino acid sequence of the antigenic epitope peptide is SEQ ID NO:3. The antigenic epitope peptide obtained by this invention exhibits good exposure, facilitating efficient antibody recognition and ensuring the reliability and efficiency of the detection reaction. An indirect ELISA detection method established using this epitope peptide as the coating antigen significantly reduces non-specific binding and background interference compared to using the full-length N protein, resulting in a substantial improvement in detection specificity. In clinical sample validation, the concordance rate reached 100%, and no cross-reactivity was observed. Furthermore, in terms of sensitivity, this method can detect positive sera at higher dilutions, with a detection limit significantly superior to that of the full-protein antigen.

The invention relates to the technical field of antibodies, and particularly provides a truncated fragment of African swine fever virus C129R protein, an antibody or an antigen binding fragment of the antibody and application of the truncated fragment and the antibody. The amino acid sequence of the truncated fragment C129R delta C is coded by SEQ ID NO: 1, and the truncated fragment C129R delta C has improved immunogenicity. The antibody specifically bound with the C129R protein or the antigen binding fragment of the antibody comprises a heavy chain variable region as shown in SEQ ID NO: 2 and a light chain variable region as shown in SEQ ID NO: 4; the blocking rate of the monoclonal antibody 3C10 is higher than 90%. On the basis of the core material, a kit for detecting the indirect ELISA antibody and the blocking ELISA antibody is constructed. The two kits both show high sensitivity, high specificity and good repeatability, and an efficient and reliable tool is provided for serological detection of African swine fever.

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.