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

Mink parvovirus enteritis pathogen antigen colloidal gold detection test strip and preparation method thereof

A mink parvovirus and toxic enteritis technology, which is applied to measuring devices, instruments, scientific instruments, etc., can solve the problems of cumbersome HA operation, high morbidity and mortality of mink viral enteritis, and no treatment method

Inactive Publication Date: 2018-06-12
INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS
View PDF2 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] The morbidity and mortality of mink viral enteritis caused by mink enteritis virus (MEV) are high, which has brought huge economic losses to the mink industry
However, there is no specific treatment for this disease, and it can only be prevented by vaccine immunization, so the diagnosis of mink viral enteritis is particularly important
At present, the diagnosis of MEV is mainly HA-HI and traditional PCR method. HA operation is cumbersome, time-consuming and laborious, and requires the preparation of pig red blood cells; while PCR is a molecular biological diagnosis method developed in recent years, which is mainly used for viruses. Nucleic acid detection, but the PCR method requires professional equipment, and operators need special training to operate in a specialized laboratory

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
  • Mink parvovirus enteritis pathogen antigen colloidal gold detection test strip and preparation method thereof
  • Mink parvovirus enteritis pathogen antigen colloidal gold detection test strip and preparation method thereof
  • Mink parvovirus enteritis pathogen antigen colloidal gold detection test strip and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0041]The preparation method of the mink parvovirus enteritis pathogen antigen colloidal gold detection test strip of the present invention mainly comprises the following steps:

[0042] Step 1, prokaryotic expression of mink parvovirus (MEV) VP2 protein and its purification

[0043] Step 2, prokaryotic expression of mink parvovirus (MEV) VP1 partial protein and its purification

[0044] Step 3, preparation of mink parvovirus (MEV) monoclonal antibody

[0045] Step 4, preparation of colloidal gold solution

[0046] Step 5. Preparation of gold-labeled antibody solution

[0047] Step 6. Preparation of gold standard pad

[0048] Step 7. Pretreatment of NC membrane (marking of C and T lines)

[0049] Step eight, assembly of test strips.

Embodiment 1

[0050] Example 1 Prokaryotic expression of mink parvovirus (MEV) VP2 protein and its purification

[0051] The VP2 protein of the detection line (T line) is obtained through prokaryotic expression and purification.

[0052] The specific process is as follows: (1) PCR technique is used to amplify mink parvovirus MEVB strain VP2 gene, the amplification conditions are: 95°C, 5min; 94°C, 30s, 55°C, 1min, 72°C, 2min; 72°C, 8min. The obtained gene fragments were directional cloned into the prokaryotic expression vector PET-30a, and the prokaryotic expression recombinant plasmid PET-30a-VP2 of mink parvovirus (MEV) VP2 gene was constructed.

[0053] (2) The prokaryotic expression recombinant plasmid PET-30a-VP2 of mink parvovirus (MEV) VP2 gene obtained above was transformed into the host strain BL21, and the VP2 protein was expressed in the form of inclusion bodies after IPTG induction, and analyzed by SDS-PAGE and Western blotting It shows that the VP2 protein can be expressed cor...

Embodiment 2

[0055] Example 2 Prokaryotic expression of mink parvovirus (MEV) VP1 partial protein and its purification

[0056] Mink parvovirus (MEV) VP1 protein is obtained by expressing part of the VP1 gene, and its expression and purification method refers to Example 1.

[0057] The specific process is as follows: (1) Use PCR technology to amplify part of the VP1 gene of mink parvovirus MEVB strain, the amplification conditions are: 95°C, 5min; 94°C, 30s, 55°C, 45s, 72°C, 90s; 72°C, 8min . The obtained gene fragments were directional cloned into the prokaryotic expression vector PET-30a, and the prokaryotic expression recombinant plasmid PET-30a-VP1 of mink parvovirus (MEV) VP1 gene was constructed.

[0058] (2) Transform the prokaryotic expression recombinant plasmid PET-30a-VP1 of the mink parvovirus (MEV) VP1 gene obtained above into the host strain BL21. After IPTG induction, the VP1 protein is expressed in the form of inclusion bodies, and analyzed by SDS-PAGE and Western blotting...

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 discloses a colloidal gold detection test strip for the pathogen antigen of mink parvovirus enteritis and a preparation method thereof, and relates to the technical field of detection of fur animal viral infectious disease pathogens. The colloidal gold detection test strip of the present invention comprises: a sample pad, a gold standard pad, an NC film and an absorption pad, the detection line of the NC film is sprayed with VP2 protein, and the quality control line of the NC film is sprayed with goat anti-mouse II anti. The preparation method of the colloidal gold detection test strip for mink parvovirus enteritis pathogen antigen of the present invention mainly comprises the following steps: prokaryotic expression of mink parvovirus (MEV) VP2 and VP1 proteins and purification thereof; mink parvovirus (MEV) monoclonal antibody preparation of colloidal gold solution; preparation of gold-labeled antibody solution; preparation of gold-labeled pad; pretreatment of NC membrane; assembly. The invention helps fur animal farms to purify mink parvoviral enteritis and facilitates the operation of grassroots veterinary personnel.

Description

technical field [0001] The invention relates to the technical field of detecting pathogens of fur animal viral infectious diseases, in particular to a colloidal gold detection test strip for mink parvovirus enteritis pathogen antigen and a preparation method thereof. Background technique [0002] The morbidity and mortality of mink viral enteritis caused by mink enteritis virus (MEV) are high, which has brought huge economic losses to the mink industry. However, there is no specific treatment for this disease, and it can only be prevented by vaccine immunization, so the diagnosis of mink viral enteritis is particularly important. At present, the diagnosis of MEV is mainly HA-HI and traditional PCR method, HA operation is cumbersome, time-consuming and laborious, and requires the preparation of pig red blood cells; and PCR is a molecular biological diagnosis method developed in recent years, mainly used for virus Nucleic acid detection, but the PCR method requires profession...

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): G01N33/577G01N33/569G01N33/558
CPCG01N33/558G01N33/56983G01N33/577G01N2333/015
Inventor 王建科程悦宁张淼程世鹏林鹏赵航易立仝明薇曹智刚孙亚茹闫喜军陈立志
Owner INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS
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