Blocking ELISA kit for detecting NDV (Newcastle disease virus) antibody
A technology for Newcastle disease virus and antibody detection, which is applied in the field of Newcastle disease virus antibody detection blocking ELISA kits, and achieves the effects of good repeatability, high-throughput detection and easy operation.
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Embodiment 1
[0027] Example 1 Preparation of Newcastle Disease Virus Antibody Detection Blocking ELISA Kit
[0028] 1. Preparation of Newcastle disease virus antigen
[0029] When chicken embryos are incubated to 9-10 days of age, get Newcastle disease virus (NDV / Chicken / Guangdong / 2008 / G7) preserved at -80°C (isolated and identified from Guangdong diseased chicken flock in 2008 by the inventor's laboratory) Dilute with PBS after thawing, take 0.1mL / piece to inoculate chicken embryos, continue to incubate at 37°C for 24 hours, then illuminate the embryos, discard dead chicken embryos, and continue to incubate at 37°C for 72 hours. Collect the allantoic fluid of chicken embryos, then centrifuge at 8000rpm for 10min at high speed to remove impurities, collect the supernatant, inactivate with β-propiolactone (0.05%) at 4°C overnight, and centrifuge the inactivated virus at 35000rpm for 3h, discard the supernatant , the pellet was resuspended in PBS, and the protein concentration was determine...
Embodiment 2
[0078] Example 2 Assembly of Newcastle disease virus antibody detection blocking ELISA kit
[0079] Newcastle disease virus antibody detection blocking ELISA kit, including: horseradish peroxidase-labeled Newcastle disease virus NP protein monoclonal antibody, ELISA plate coated with Newcastle disease virus inactivated antigen, Newcastle disease virus positive control serum, Newcastle disease virus Epidemic virus negative control serum, serum diluent, wash solution, substrate solution and stop solution.
[0080] Horseradish peroxidase-labeled Newcastle disease virus NP protein monoclonal antibody: secreted by a cell line with the deposit number CCTCC NO: C2016180, and then purified;
[0081] ELISA plate coated with inactivated Newcastle disease virus antigen: Dilute the concentration of inactivated Newcastle disease virus antigen to 1ng / μL, coat the ELISA plate with 100μL / well, incubate at 37°C for 2 hours or overnight at 4°C;
[0082] Positive control serum: chicken serum af...
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