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.

Active Publication Date: 2017-04-26
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The current ELISA method for Newcastle disease virus antibody detection is indirect ELISA, which can only be used for chicken or turkey serum samples

Method used

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  • Blocking ELISA kit for detecting NDV (Newcastle disease virus) antibody
  • Blocking ELISA kit for detecting NDV (Newcastle disease virus) antibody
  • Blocking ELISA kit for detecting NDV (Newcastle disease virus) antibody

Examples

Experimental program
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Effect test

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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Abstract

The invention discloses a blocking ELISA kit for detecting an NDV (Newcastle disease virus) antibody. The blocking ELISA kit for detecting the NDV antibody comprises an ELISA plate coated with an NDV inactivated antigen, an NDV positive control serum, an NDV negative control serum and a horseradish peroxidase labeled NDV NP protein monoclonal antibody, wherein the horseradish peroxidase labeled NDV NP protein monoclonal antibody is secreted by a hybridoma cell strain with the preservation number being CCTCC NO: C2016180. The blocking ELISA kit for detecting the NDV antibody can detect serum samples which are infected with the suspected NDV and are from different species, can distinguish an MG7-deficient vaccine from an NDV serum after being infected with a wild virus, and has no cross reaction with a common avian viral pathogen positive serum, thereby being high in sensitivity and specificity, good in reproducibility and suitable for high-throughput detection of serum samples.

Description

technical field [0001] The invention relates to a kit for veterinary detection, in particular to an ELISA kit for detecting and blocking Newcastle disease virus antibody, and belongs to the field of preparation of the ELISA kit for detecting and blocking Newcastle disease virus antibody. Background technique [0002] Newcastle Disease (ND) is a highly contagious disease of poultry caused by Newcastle Disease Virus (NDV). Huge economic loss seriously affects the development of poultry industry. [0003] NDV belongs to the genus Mumpsvirus of the Paramyxoviridae family, has an envelope structure, and its genome is a single-strand negative-sense RNA virus that is not segmented. It is composed of 6 structural proteins, namely NP, P, M, F, HN and L. protein. NP protein is a nucleocapsid protein with high amino acid conservation. Most NDV viruses have a highly conserved B cell epitope at position 443-460 of NP protein; P protein is an essential factor for viral RNA synthesis; M,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/531
CPCG01N33/531G01N33/56983G01N2333/125
Inventor 朱启运贾艳娥周广青吉艳红杨麦娟
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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