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

Bi-combined blocking ELISA antibody detection kit for detecting porcine PrV (pseudorabies virus) and FMDV (foot and mouth disease virus) and application of kit

A technology for porcine pseudorabies virus and foot-and-mouth disease virus, which is applied in the field of animal disease detection, can solve problems such as increased losses, lag in disease prevention and control measures, and excessively long detection time, and achieves shortened detection time, reduced sample serum volume, and good specificity. Effect

Inactive Publication Date: 2018-05-04
WUHAN KEQIAN BIOLOGY CO LTD
View PDF9 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Differential diagnosis is to find problems in the early stage of the epidemic and take timely prevention and control measures to reduce losses. If the detection time is too long, the disease prevention and control measures will undoubtedly lag behind and lead to the expansion of losses.

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
  • Bi-combined blocking ELISA antibody detection kit for detecting porcine PrV (pseudorabies virus) and FMDV (foot and mouth disease virus) and application of kit
  • Bi-combined blocking ELISA antibody detection kit for detecting porcine PrV (pseudorabies virus) and FMDV (foot and mouth disease virus) and application of kit
  • Bi-combined blocking ELISA antibody detection kit for detecting porcine PrV (pseudorabies virus) and FMDV (foot and mouth disease virus) and application of kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] The preparation of embodiment 1 antigen-coated plate

[0038] The preparation of the antigen detection plate that is coated with porcine pseudorabies virus and foot-and-mouth disease virus 3B polypeptide comprises the steps:

[0039] (1) Porcine pseudorabies virus cultivation and purification: the porcine pseudorabies virus (E A strain) virus was inoculated into BHK cells with a 5% ratio, and the virus liquid was collected after 48 hours; the collected virus liquid was concentrated for 30 days through a hollow fiber with a pore size of 100KD times, and then the concentrated solution was purified by Sephadex molecular sieves, and the virus solution corresponding to the characteristic peak was collected as the final porcine pseudorabies virus antigen solution, which was stored below -15°C for later use;

[0040] (2) Send the 3B polypeptide sequence in the 3ABC protein to the protein synthesis company through the published O-type foot-and-mouth disease protein sequence, an...

Embodiment 2

[0045] Preparation of embodiment 2 enzyme-labeled monoclonal antibody

[0046] (1) Preparation of anti-pseudorabies virus gE protein monoclonal antibody: BALB / c mice were immunized with purified porcine pseudorabies virus (E A strain), and the splenocytes and mouse myeloma cells (SP2 / 0) were used for Cell fusion, after culturing with HAT selective medium, then use purified porcine pseudorabies virus gE protein-coated plate for indirect ELISA screening, the screened positive cell lines are cloned by limiting dilution method, and then porcine pseudorabies virus (Hubei A strain, gE gene is not deleted) positive control serum of pigs infected with porcine pseudorabies virus and negative serum of healthy pigs not infected with porcine pseudorabies virus were screened by blocking ELISA, and the monoclonal antibody with blocking effect was finally screened, and the secretion of this monoclonal antibody The cell line was named PRV-eA-gE.

[0047] (2) Preparation of anti-foot-and-mout...

Embodiment 3

[0054] Example 3 kit assembly

[0055] The kit contains 2 pieces of 96-well ELISA antigen detection plates, 1 bottle of mixed solution of two enzyme-labeled monoclonal antibodies (20mL / bottle), 1 bottle of porcine pseudorabies virus gE protease-labeled monoclonal antibody chromogenic substrate solution, and 1 bottle of stop solution (10mL / bottle), 1 bottle of foot-and-mouth disease virus 3ABC protease-labeled monoclonal antibody chromogenic substrate solution and stop solution (10mL / bottle), 2 tubes of negative control and positive control (1.5mL / tube), 1 bottle of sample diluent (50mL / bottle) and 1 bottle of concentrated washing solution (30mL / bottle), with an instruction manual attached.

[0056] The above-mentioned antigen detection plate is an antigen detection plate coated with porcine pseudorabies virus and foot-and-mouth disease virus 3B polypeptide;

[0057] The above two enzyme-labeled monoclonal antibodies are horseradish peroxidase-labeled anti-pseudorabies virus g...

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

No PUM Login to View More

Abstract

The invention belongs to the field of animal epidemic disease detection and particularly relates to a bi-combined blocking ELISA antibody detection kit for detecting a porcine PrV (pseudorabies virus)gE protein antibody and an FMDV (foot and mouth disease virus) 3ABC protein antibody and an application of the kit. The kit comprises an antigen detection plate coated with porcine PrV and FMDV 3B polypeptides, a mixed solution containing an enzyme-labeled anti-PrV gE protein monoclonal antibody and an enzyme-labeled anti-FMDV 3ABC protein monoclonal antibody, a porcine PrV gE protein enzyme-labeled monoclonal antibody developing system, a FMDV 3ABC protein enzyme-labeled monoclonal antibody developing system, a negative / positive control, a sample diluent and a washing solution. Two independent ELISA reactions are performed simultaneously in first two steps, unique developing systems are used, so that developing and termination of the last enzyme do not affect the reaction of the next enzyme, one sample incubation and one second-antibody incubation are realized for one part of serum, and two ELISA results are obtained.

Description

technical field [0001] The invention belongs to the field of animal disease detection, and in particular relates to a dual blocking ELISA antibody detection kit for detecting porcine pseudorabies virus gE protein antibody and foot-and-mouth disease virus 3ABC protein antibody and application thereof. Background technique [0002] Pseudorabies (PR) is an important infectious disease caused by pseudorabies virus (Pseudorabies Virus, PrV), including domestic animals and various wild animals, with symptoms of fever, itching and encephalomyelitis. The disease has caused huge economic losses to the pig industry, and pigs are the natural host and storage of the disease. Prevention, control and ultimate eradication of this disease is a difficult task that our country is currently facing. [0003] In the prevention and control of pseudorabies, gE gene-deleted live vaccines are generally used for immunization. These vaccines do not produce antibodies against gE protein after immuniza...

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 Applications(China)
IPC IPC(8): G01N33/569G01N33/577
CPCG01N33/56983G01N33/577
Inventor 陈斌但汉并徐高原尹争艳陈关平
Owner WUHAN KEQIAN BIOLOGY CO LTD
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