Double-antibody sandwich method for quickly detecting novel H1N1 influenza virus

A double-antibody sandwich and influenza virus technology, which is applied in the direction of introducing foreign genetic material, application, and measuring devices by using carriers, can solve the problems of high cost, long time for isolation and identification of influenza virus, and difficult promotion and application at border ports, etc., to achieve effective antigenicity Diagnosis, easy to use, accurate detection effect

Inactive Publication Date: 2011-06-15
CHINESE ACAD OF INSPECTION & QUARANTINE
View PDF2 Cites 6 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the isolation and identification of influenza virus takes a long time and costs high, and needs to be carried out in biosafety laboratories or national designated research institutions, while RT-PCR...

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
  • Double-antibody sandwich method for quickly detecting novel H1N1 influenza virus
  • Double-antibody sandwich method for quickly detecting novel H1N1 influenza virus
  • Double-antibody sandwich method for quickly detecting novel H1N1 influenza virus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0028] The present invention is a double-antibody sandwich method for rapid detection of novel H1N1 influenza virus. The key technology lies in: using genetic engineering method to prepare novel H1N1 influenza virus hemagglutinin protein HA1 N-terminal subunit-specific recombinant antigen, and immunizing with the antigen Rabbit, the prepared rabbit serum was used for ELISA coating, and the rabbit anti-H1N1 hemagglutinin protein labeled with horseradish peroxidase was used as the detection antibody, and a double-antibody sandwich detection method for the new H1N1 influenza virus was established.

[0029] Technical route of the present invention is as follows:

[0030] Design and amplify the expression of the new H1N1 influenza virus HA1 N-terminal spherical subunit primer→PCR amplification→ligate the target fragment with the vector pET100 / D-TOPO, construct the recombinant plasmid→transform the expression cell (Escherichia coli BL21)→IPTG induce the expression of the recombinant ...

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
Concentrationaaaaaaaaaa
Login to view more

Abstract

The invention discloses a double-antibody sandwich method for quickly detecting a novel H1N1 influenza virus, comprising the following specific steps: designing the HA1N end spherical sub-gene primer of an amplified-expression novel H1N1 influenza virus; carrying out PCR (Polymerase Chain Reaction) amplification; connecting a target segment with a carrier pET100/D-TOPO (trioctyl-phosphine oxide), and building a recombinant plasmid; converting an expression cell (escherichia coli BL21); expressing IPTG (isopropyl beta-D-thiogalactopyranoside) induced recombinant protein; analyzing the antigenicity of a purified product by indirect ELISA (enzyme-linked immuno sorbent assay) and Western blottin; purifying the recombinant protein by a HisTrapTMHP (histidine) affinity adsorption column purification system; preparing rabbit serum by a recombinant protein immune rabbit; carrying out ELISA coating by the prepared rabbit serum; taking the rabbit resisting H1N1 influenza virus hemagglutinin protein marked by a horse radish peroxidise as a second antibody; and building a double-antibody sandwich method for detecting the novel H1N1 influenza virus antigen. The method can be used for carrying out quick and effective antigen diagnosis, is convenient to use and accurate to detect, and is suitable for being popularized and applied to areas, such as border ports and the like.

Description

technical field [0001] The invention relates to a double-antibody sandwich method for rapid detection of novel H1N1 influenza virus. Background technique [0002] Influenza A (H1N1 Influenza A (H1N1) virus) is a highly contagious respiratory virus that has been listed as a weapon of bioterrorism. The virus of the 2009 influenza pandemic was a new type of H1N1 virus. This influenza caused a worldwide pandemic and caused great harm to human health. It is imminent to establish a rapid detection method to prevent the occurrence of influenza disease. At present, the methods for detecting the novel H1N1 influenza virus mainly include virus isolation and culture, reverse transcription polymerase chain reaction (RT-PCR), serological diagnosis and antigen detection. However, the isolation and identification of influenza virus takes a long time and costs high, and needs to be carried out in biosafety laboratories or national designated research institutions, while RT-PCR relies on sp...

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): G01N33/569C12N15/44C12N15/63C07K16/18
Inventor 杨宇胡健萍王静
Owner CHINESE ACAD OF INSPECTION & QUARANTINE
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap