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Kit for synchronously detecting African swine fever virus and porcine pseudorabies virus

A technology for African swine fever virus and porcine pseudorabies virus, which can be used in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., and can solve the problems of high detection cost, time-consuming and laborious

Active Publication Date: 2021-11-05
WUHAN INST OF VIROLOGY CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Commercialized PCR kits are mostly single-item PCR. One kit can only detect one pathogen in one test. For the same swine disease material mixed with two or more pathogens, two or more pathogens must be diagnosed. The above kits and two or more PCR tests are not only time-consuming and labor-intensive, but also costly.

Method used

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  • Kit for synchronously detecting African swine fever virus and porcine pseudorabies virus
  • Kit for synchronously detecting African swine fever virus and porcine pseudorabies virus
  • Kit for synchronously detecting African swine fever virus and porcine pseudorabies virus

Examples

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Embodiment 1

[0029] In order to detect pathogenic bacteria in the production of multi - disease mixed infections in pig production, the inventors designed a kit for synchronizing an African swine fever virus and psessy rabies virus, the kit for multiple fluorescent quantitative PCR kits, against Africa The swine fever virus CD2V gene, the African swine fever virus P72 gene and the porchiar rabies virus EPO gene were detected. This kit includes primer pairs for amplifying African swine fever CD2V genes, African swine fever P72 genes and pseudo rabies virus EPO genes, and corresponding fluorescent probes, according to the ASFV CD2V, ASFV-P72, and ASFV-P72, and The gene sequence of PRV-EPO is designed to amplify primers for amplification of CD2V genes on ASFV-CD2V-F and ASFV-CD2V-R and their corresponding fluorescent probes ASFV-CD2V-P for amplifying the primer pair of P72 genes. ASFV-P72-F and ASFV-P72-R and corresponding fluorescent probes ASFV-P72-P for amplifying the primer of EPO pair is PRV...

Embodiment 2

[0044] Example 2 A method of synchronizing an African swine fever virus and a pseudo rabies virus

[0045] The detection of African swine fever viruses and psessic rabies viruses were performed using synchronous detection of African swine fever viruses and pocalyptic rabies viruses using synchronization in Example 1.

[0046] According to the African Pine Virus (ASFV), the nucleic acid sequences of the Pig Pseudo-rabies virus (PRV), and the above-mentioned three plasmids are linearized using the above three plasmids. Treatment, after the gel electrophoresis is recovered, the concentration of these three DNA fragments that are collected by the spectrophotometer is measured, and the copy number of these three DNA fragments is obtained according to the general copy number conversion formula. Dilute DNA fragment into 1x10 according to copy number gradient 6 Copies / UL, 1x10 5 Copies / UL, 1x10 4 Copies / UL, 1x10 3 Copies / UL, 1x10 2 Copies / UL, 1x10 1 Copies / UL, 1x10 0 Copies / ...

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Abstract

The invention relates to a kit for synchronously detecting African swine fever virus and porcine pseudorabies virus. The kit is a multiplex fluorescent quantitative PCR kit, and comprises primer pairs for amplifying an African swine fever virus CD2v gene, an African swine fever virus P72 gene and a porcine pseudorabies virus EPO gene, and corresponding fluorescent probes, wherein the nucleotide sequences of the primer pair for amplifying the CD2v gene and the corresponding fluorescent probe thereof, the primer pair for amplifying the P72 gene and the corresponding fluorescent probe thereof, and the primer pair for amplifying the EPO and the corresponding fluorescent probe thereof are as shown in SEQ ID NO: 1-9 correspondingly.

Description

Technical field [0001] The present invention relates to the field of gene detection, and more particularly to a kit for synchronizing an African swine fever virus and a pseudo rabies virus. Background technique [0002] In recent years, a variety of pathogen mixed infections in pig production have become common. The disease is not caused by a single pathogens, but caused by two or more pathogens, delayed diagnosis and control, leading to high incidence and high mortality rate of pigs, causing great economic losses. In the presence of multi-disease mixed-infected pig farms, pig diseases are complicated, and the case changes are not typical. The condition is serious, and the site is difficult to diagnose, and general prevention measures are also difficult to work. The detection of each virus can be used to use traditional diagnostic methods, such as virus separation detection or serological detection methods. There is a cumbersome method, the diagnosis time and the problem of the p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/6851C12N15/11
CPCC12Q1/701C12Q1/6851C12Q2600/16C12Q2600/166C12Q2531/113C12Q2537/143C12Q2563/107C12Q2545/113Y02A50/30
Inventor 苏正元王华林邓菲杨娟
Owner WUHAN INST OF VIROLOGY CHINESE ACADEMY OF SCI
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