Fluorescence quantitative PCR detection reagent for Asf virus and preparation method and use thereof

An African swine fever virus and fluorescence quantitative technology, which is applied in the field of biological preparations, can solve the problems that the rapid detection method of African swine fever virus has not been carried out, the detection reagents of the African swine fever virus, the animal inoculation test is not safe, and the virus cannot be directly detected, etc., reaching the scope of application of the sample Wide, intuitive results, easy to operate

Inactive Publication Date: 2006-10-04
CHECKOUT & QUARANTINE TECH CENT YUNNAN ENTRY &EXIT CHECKOUT & QUARANTINE BUR
View PDF0 Cites 12 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, the sensitivity and specificity of red blood cell adsorption test and direct immunofluorescence test are not high, and the animal inoculation test is not safe. ELISA has great practicability in quarantine and diagnosis, and is sensitive, specific, and the results are highly consistent. characteristics, but the ELISA method can only detect antibodies and cannot directly detect viruses
The polymerase chain reaction (PCR) method for detecting ASFV has the characteristics of specificity, sensitivity, and rapidity, but it has disadvantages such as time-consuming, easy to pollute, electrophoresis detection after amplification, and a small number of samples for each detection.
So far, there has been no report on the rapid detection method of African swine fever virus and related detection reagents in China.

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
  • Fluorescence quantitative PCR detection reagent for Asf virus and preparation method and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0030] 1. Construction of ASFV VP73 Gene Recombination Plasmid

[0031] According to "Molecular Cloning Experiment Guide" (Third Edition) translated by Huang Peitang et al., China Science Press, January 2003, page 1217 to page 1259, African swine fever virus VP73 gene was cloned and sequenced, and ASFV VP73 gene recombination was constructed Plasmid, the recombinant plasmid was named pBAD-VP73.

[0032] 2. Design primers and probes

[0033] The present invention selects the conserved fragment of the ASFV VP73 gene as the target, and performs homology analysis and comparison on the DNA sequence of the ASFV gene reported in GenBank and the DNA sequence of the African swine fever virus gene cloned and sequenced in 1 above, and selects the ASFV gene. The most conserved fragment (66bp), using primer Express software and primer prere5.0 software, design primers and probes, the synthesis of primers and probes adopts β-acetonitrile phosphoramidite chemical synthesis method, and uses ...

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 related bio-reagent is designed by: with conservative African swine fever virus VP73 gene fragment as target object, designing and composing carefully for large quantities of primers and probes, taking optimal pairing screen test on different conditions to obtain the most proper primer and probe. The fluorescence quantitative RT-PCR testing reagent includes one couple of specific primers and one specific fluorescent probe, and the amplification fragment length is 66bp. This invention is fit to fast testing for African swine fever.

Description

technical field [0001] The invention relates to a biological agent designed and synthesized by taking the gene fragment of African swine fever virus as a target, especially a reagent capable of detecting the African swine fever virus, and a preparation method and application of the reagent. Background technique [0002] African swine fever (ASF) is a contagious viral disease of pigs that attacks macrophages and monocytes and is typically characterized by high mortality, reddened areas of the skin and severe bleeding in internal organs. In countries and regions where the disease occurs, the mortality rate can be as high as 100%. Under natural conditions, ASFV is lodged in Ornithodoros ticks and recessively infected African wild boars, warthogs, South African wild boars and giant wild boars. So far only pigs have been found to show clinical signs. [0003] After African swine fever was first discovered in Kenya in East Africa in 1909 in domestic pigs brought by European immi...

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): C12Q1/68G01N21/64
Inventor 花群义周晓黎董俊杨云庆徐自忠肖荣海尹尚莲
Owner CHECKOUT & QUARANTINE TECH CENT YUNNAN ENTRY &EXIT CHECKOUT & QUARANTINE BUR
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