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

H7 subtype avian influenza virus monoclonal antibody of and kit

An avian influenza virus, monoclonal antibody technology, applied in antiviral immunoglobulin, biochemical equipment and methods, instruments, etc., can solve the problems such as no monoclonal antibody yet, achieve easy popularization, strong sensitivity, specificity good effect

Active Publication Date: 2013-08-28
哈尔滨元亨生物药业有限公司
View PDF3 Cites 16 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] But so far, there are no reports of monoclonal antibodies and corresponding detection kits for H7 subtype avian influenza virus with good sensitivity, strong specificity and high titer

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
  • H7 subtype avian influenza virus monoclonal antibody of and kit
  • H7 subtype avian influenza virus monoclonal antibody of and kit
  • H7 subtype avian influenza virus monoclonal antibody of and kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Embodiment 1, the acquisition of H7 subtype avian influenza virus HA gene monoclonal antibody hybridoma cell

[0056] 1. Propagation and purification of H7N2 avian influenza virus

[0057] H7N2 avian influenza virus was inoculated into 9-day-old fertilized SPF chicken embryos, and after incubation at 30°C for 2 days, the allantoic fluid of the chicken embryos was collected to obtain the amplified H7N2 strain virus. Collect the H7N2 avian influenza virus and inactivate it with 0.03% formalin at 4°C. The inactivated H7N2 avian influenza virus is detected by HA to determine the titer of the inactivated H7N2 avian influenza virus solution (the details of HA titer determination and HI detection Refer to WHO for the method, we choose HA=1024, the H7N2 avian influenza virus strain is provided by the China Animal Disease Prevention and Control Center).

[0058] Centrifuge the H7N2 avian influenza virus inactivated virus liquid at 8000rpm for 30 minutes, keep the supernatant, l...

Embodiment 2

[0084] Example 2, H7 Subtype Avian Influenza Virus HA Antigen ELISA Kit (Enzyme-Linked Immunoassay, ELISA)

[0085] 1. Preparation of ELISA plate:

[0086] Pre-coat the rabbit polyclonal antibody against H7N2 avian influenza virus on the polyethylene microwell plate of the kit. The polyclonal antibody is coated with 10mM carbonate buffer (pH9.6) at 4°C Overnight; then wash twice with PBST (10mM PBS+0.05%Tween20), then add blocking solution (10mM PBS+2% bovine serum albumin, BSA), block at 37°C for 2 hours, dry and vacuum-dry and pack into a finished kit ELISA plate.

[0087] 2. Configuration of other components of the kit:

[0088] A) The composition of virus lysate:

[0089] Lysis buffer A (LB-A): 6%CHAPS+2%Tween-20+1%Tween-80.

[0090] Lysis solution B (LB-B): 100mM PMSF, dissolved in isopropanol, the final working concentration is 2mM.

[0091] Lysis solution C (LB-C): 10mM PBS, pH7.4

[0092] B) Enzyme-labeled reagents:

[0093] The enzyme-labeled monoclonal antibod...

Embodiment 3

[0127] Example 3, H7 Subtype Avian Influenza Virus Antibody ELISA Kit (Enzyme-Linked Immunoassay, ELISA)

[0128] 1. Preparation of ELISA plate:

[0129] Pre-coat the H7N2 avian influenza virus purified antigen on the polyethylene microwell plate of the kit, use 10mM carbonate buffer (pH9.6) as the coating solution, and coat overnight at 4°C; then use PBST (10mM PBS +0.05%Tween20) wash twice, then add blocking solution (10mM PBS+2% bovine serum albumin BSA), block at 37°C for 2 hours, remove the liquid and dry in the air, after drying, vacuum pack into a finished kit enzyme target board.

[0130] 3. Configuration of other components of the kit:

[0131] A) Enzyme-labeled reagent: prepare enzyme-labeled monoclonal antibody by sodium periodate oxidation method, the specific steps are as follows. Weigh 5mg of HRP, dissolve it in 1ml of pure water, the solution turns brown; add 1ml of freshly prepared 0.06mol / L NaIO4 solution (12.8mg / ml), stir in a molecular hybridization oven ...

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 provides an H7 subtype avian influenza virus monoclonal antibody of and kit, and obtains hybridoma cell strain that secretes H7 subtype avian influenza virus monoclonal antibody, wherein the accession number of the hybridoma cell strain is CGMCC NO. 7308, and the obtained H7 subtype avian influenza virus monoclonal antibody is provided with advantages like high sensitivity, strong singularity and high bioactivity etc., which can be used to identify if the virus to be tested is H7 subtype avian influenza virus, and if the sample to be tested is infected by H7 subtype avian influenza virus, and if the serum to be tested contains H7 subtype avian influenza virus. The monoclonal antibody above also could be used to prepare immunological diagnostic reagent of H7 subtype avian influenza virus or its antibody, such as Enzyme-linked diagnostic reagent and colloidal gold immunoassay kit or the like, and also could detect if the poultry feces, the oral and nasal secreta or the stool contains H7 subtype avian influenza virus and if the poultry serum contains H7 subtype avian influenza virus antibody.

Description

technical field [0001] The invention belongs to biological detection technology, and relates to a monoclonal antibody of H7 subtype avian influenza virus and a kit. Background technique [0002] Avian influenza (Avian Influenza, AI), referred to as avian influenza, is an infectious disease of poultry caused by influenza A virus of the Orthomyxoviridae family. Avian Influenza virus (AIV) can infect almost all wild birds and poultry. According to the different pathogenicity of AIV, it can be divided into highly pathogenic, low pathogenic and non-pathogenic AIV. In recent years, avian influenza has become prevalent worldwide, causing huge economic losses to the poultry industry. Avian influenza virus is very easy to mutate and can spread across species to infect humans, becoming one of the important pathogens that threaten human health. In Hong Kong in 1997, in the Netherlands in 2003, and in Southeast Asia since 2004, there have been cases of death due to human infection wit...

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
IPC IPC(8): C12N5/20C07K16/10G01N33/577C12R1/91
Inventor 陈西钊孙明张丽马永缨迟立超秦亚嫚
Owner 哈尔滨元亨生物药业有限公司
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