Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Competitive ELISA antibody detection kit for bovine infectious rhinotracheitis virus and its application

A rhinotracheitis virus and antibody detection technology is applied in the field of antibody detection of bovine infectious rhinotracheitis virus, which can solve the problem of not seeing bovine infectious rhinotracheitis virus, etc., and achieve good consistency and sensitivity of test results. Good, high sensitivity effect

Active Publication Date: 2021-11-26
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no relevant report on the competitive ELISA antibody detection method for bovine infectious rhinotracheitis virus

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Competitive ELISA antibody detection kit for bovine infectious rhinotracheitis virus and its application
  • Competitive ELISA antibody detection kit for bovine infectious rhinotracheitis virus and its application
  • Competitive ELISA antibody detection kit for bovine infectious rhinotracheitis virus and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Embodiment 1. Design and expression of IBRV recombinant gB protein

[0058] 1. Sequence optimization and primer synthesis

[0059] Download the IBRV gB gene sequence (GeneID: 4783419) from the GenBank database, use the online analysis website (http: / / www.cbs.dtu.dk / services / TMHMM / ) to analyze the sequence, and design a truncated gB gene sequence ( SEQ ID NO: 1), which encodes a truncated gB protein (SEQ ID NO: 2). In order to secrete and express the truncated gB protein in insect cells, the codon optimization of the truncated gB gene sequence was performed according to insect cell preference to obtain the target gene sequence (SEQ ID NO: 3). Entrust Sangon Bioengineering (Shanghai) Co., Ltd. to synthesize the target gene. According to the base sequence of the baculovirus expression vector and the target gene sequence, a pair of homology arm amplification primers were designed, and restriction sites NdeI (CATATG) and XhoI (CTCGAG) were added to the upstream and downstr...

Embodiment 2

[0079] Example 2. Preparation and screening of anti-IBRV monoclonal antibodies

[0080] 1. Animal Immunization

[0081] First immunization: After mixing the purified IBRV virus liquid (Bartha Nu / 67 standard strain) with an equal volume of Freund's complete adjuvant (Sigma-Aldrich, product number F5881) and emulsifying completely, immunize 6-8 week-old SPF grade female BALB / c mice, the immunization dose is 50 μg / mouse. Two weeks after the first immunization, an equal volume of IBRV virus fluid (Bartha Nu / 67 standard strain) was fully emulsified with Freund's incomplete adjuvant (Sigma-Aldrich, product number F5506), and the second, third and third times were carried out. 4 times of immunization, the immunization dose was 50μg / monkey, and the interval between each immunization was 2 weeks.

[0082] 2. Serum titer determination

[0083] The indirect ELISA method for hybridoma screening was established by using the positive serum of the mice after three immunizations, and the ...

Embodiment 3

[0107] Example 3. Establishment of IBRV competition ELISA antibody detection method

[0108] 1. Horseradish peroxidase-labeled monoclonal antibody

[0109] The 1F24 monoclonal antibody was labeled with horseradish peroxidase (HRP) by the sodium periodate method, and the steps were as follows:

[0110] (1) Weigh 5 mg of HRP (Sigma-Aldrich, SRE0082) and dissolve it in 1000 μL of double-distilled water, and continue to add 500 μL of newly prepared 0.1M NaIO 4 , mix well, and incubate at 4°C in the dark for 30min.

[0111] (2) Add 500 μL of freshly prepared 0.2M ethylene glycol (Sigma-Aldrich, 324558) solution, mix well, and incubate at 4°C in the dark for 30 minutes.

[0112] (3) Add 5 mg of purified 1F24 monoclonal antibody to the above solution, mix well and put it into a pre-treated dialysis bag (Solarbio, MW14000), dialyze overnight in 0.1M carbonate buffer solution with pH 9.6, and dialyze The medium was changed three times during this period.

[0113] (4) Pour the dialy...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
Dilution degreeaaaaaaaaaa
Dilution degreeaaaaaaaaaa
Dilution degreeaaaaaaaaaa
Login to View More

Abstract

The invention relates to antibody detection of bovine infectious rhinotracheitis virus, in particular to a competitive ELISA antibody detection kit of bovine infectious rhinotracheitis virus and application thereof. The invention provides a monoclonal antibody against bovine infectious rhinotracheitis virus, which is secreted by the hybridoma cell with the preservation number of CGMCC No.23005. Also provided is a bovine infectious rhinotracheitis virus antibody detection kit, which comprises the monoclonal antibody. The present invention establishes a bovine infectious rhinotracheitis virus antibody detection method based on the monoclonal antibody and the truncated bovine infectious rhinotracheitis virus gB protein designed and expressed in the present invention. The method has the advantages of strong specificity, good sensitivity, simple operation and rapidity.

Description

technical field [0001] The invention relates to antibody detection of bovine infectious rhinotracheitis virus, in particular to a competitive ELISA antibody detection kit of bovine infectious rhinotracheitis virus and application thereof. Background technique [0002] Infectious bovine rhinotracheitis virus (IBRV), also known as bovine herpesvirus-1 (BHV-1), belongs to the herpesviridae family and is an important pathogen of cattle. Infectious Bovine Rhinotracheitis (IBR). Bovine infectious rhinotracheitis is a febrile, acute and highly latent infectious disease. Since Miller first reported the disease in the United States in 1995, IBR has shown a global epidemic trend, which has caused huge economic losses to the world cattle industry. The main hazard of IBRV is that after the virus invades the host, it can latently infect the ganglion of the host, causing the sick cattle to be infected for life, which brings a huge challenge to the prevention and control of the disease. ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/08C12N5/20G01N33/577G01N33/569G01N33/68G01N33/58G01N33/543
CPCC07K16/085G01N33/577G01N33/56994G01N33/6854G01N33/581G01N33/54393C07K2317/76G01N2469/20
Inventor 刘文晓李永清程晶江波江岳王晓玥段景龙
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com