Porcine pseudorabies virus strain and application thereof

A technology for porcine pseudorabies and porcine herpes virus, which is applied in the directions of viruses, antiviral agents, virus antigen components, etc., can solve the problems of death and cannot fully protect PRV wild strains, etc., and achieves good safety, is conducive to purification, and is effective. good effect

Active Publication Date: 2020-10-09
WUHAN CHOPPER BIOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In 2011, pseudorabies broke out again. According to relevant scholars' reports, this outbreak was caused by the mutation of pseudorabies virus; according to scholars' research, the newly emerged mutant strain of PRV has different antigenicity from the classic strain, which can cause 35 Pigs over the age of one day developed neurological symptoms and died, and the Barthr-K61 strain vaccine currently on the market cannot completely protect these PRV wild strains isolated after 2011

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  • Porcine pseudorabies virus strain and application thereof
  • Porcine pseudorabies virus strain and application thereof
  • Porcine pseudorabies virus strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1: Isolation and identification of porcine pseudorabies virus

[0021] 1 separation

[0022] 1.1 Disease treatment Since 2011, swine pseudorabies epidemic has occurred in North China, East China, Central China, South China and other regions, causing sows to abort, stillbirth, mummified fetuses, and large numbers of deaths of piglets and nursery pigs. In September 2013, swine pseudorabies occurred in a pig farm in Wuhan that had been immunized with a live vaccine of Bartha-K61 strain of pseudorabies, and sows suffered abortions; suckling pigs and nursery pigs died in large numbers. We collected the brain and lung tissues of the nursery pigs that died of illness on the farm, mixed and shredded, added 5 times the volume of DMEM medium, ground in a sterilized mortar to prepare a tissue suspension, and put the tissue suspension in- After repeated freezing and thawing at 20°C for 3 times, centrifuge at 3000r / min for 30 minutes, take the supernatant and filter it with a 0....

Embodiment 2

[0030] Example 2: Genetic analysis of porcine pseudorabies virus

[0031] Variation and evolution analysis of PRV WH-13 strain: Using the designed full-gene amplification primers of gC and gD, the full lengths of gC and gD genes of PRV WH-13 strain were amplified respectively. After sequencing, they were compared with other GenBanK virus strains. The sequences are aligned and the corresponding genetic evolutionary tree is drawn.

[0032] The results showed that the length of the ORF of the gC gene of the WH-13 strain was 1464 bp, encoding 487 amino acids, and the sequencing number was NO.1. Based on the PRV gC ORF full-length gene sequence to draw a genetic evolutionary tree, such as Figure 4 . The results show that the WH-13 strain is closely related to the epidemic strains (HeN1.seq, HNX.seq, ZJ01.seq) isolated after 2011 in China, but it is closely related to the Bartha strain and the domestic strains isolated in earlier years (Ea .seq, Fa.seq) are far apart.

[0033] The ORF ...

Embodiment 3

[0034] Example 3: Virulence of porcine pseudorabies virus

[0035] 1. Toxicity test on mice:

[0036] The isolated and cultured WH-13 strain F5 virus suspension was inoculated into 5 Kunming mice, each of which was injected with 0.1 ml into the abdominal cavity, and 5 healthy mice were set as blank controls. The mice showed restlessness and screaming 24 hours after the inoculation. The coat appeared messy at 36 hours after the inoculation. They gnawed at the injection site by themselves. 5 mice died within 72-96 hours of the challenge. The control mice were healthy. live.

[0037] 2. Toxicity test on rabbits

[0038] The isolated and cultured WH-13 strain F5 generation virus suspension was inoculated with 5 healthy rabbits, each injected 1ml into the leg muscle, and 5 healthy rabbits were set as a blank control. 24 hours after the inoculation, the rabbit showed itchy, biting the injection site with the mouth, causing the injection site to lose hair, the skin was damaged, red muscles...

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Abstract

The invention discloses a porcine pseudorabies virus strain and an application thereof. The porcine pseudorabies virus WH-13 strain provided by the invention is separated in Wuhan pig farm in Hubei Province in 2013, and the identification shows that the strain is a prevalent strain with stronger virulence. The inactivated vaccine is prepared from the porcine pseudorabies virus WH-13 strain, piglets are immunized once (2ml), the serum neutralizing antibody level is not lower than 1: 128, and a protection period is as long as 6 months. The TK, gE and gI genes are deleted from the strain by adopting a genetic engineering method; the minimum immunization dose of the prepared porcine pseudorabies live vaccine piglet is 103.0 TCID50, the serum neutralizing antibody level of the piglet after immunization once is not lower than 1: 145, the immune protection rate of a classic strain and an epidemic strain is 100%, and the protection period is as long as 6 months. The preservation number of theporcine pseudorabies virus WH-13 strain is CCTCC NO: V202032.

Description

Technical field [0001] The invention belongs to the field of veterinary biological products, and specifically relates to a porcine pseudorabies virus strain and its application. Preparation method of swine pseudorabies inactivated vaccine and swine pseudorabies live attenuated vaccine. Background technique [0002] Pseudorabies virus (Pseudorabies Virus, PRV) belongs to the subfamily Herpesvirus A subfamily of the Herpesvirus family, and belongs to the varicella virus. Porcine pseudorabies (PR) is a highly contagious infectious disease caused by pseudorabies virus (PRV). The main symptoms of pigs include fever, encephalomyelitis and reproductive disorders. The disease was first discovered in the United States in 1813, and subsequently occurred in many pig-raising countries, mainly in areas with dense pig herds such as South America, Asia and Europe. It has now been distributed worldwide, and its prevalence is increasing. The Health Organization (OIE) is included in the OIE liste...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00A61K39/245A61P31/22
CPCC12N7/00A61K39/12A61P31/22C12N2710/16721C12N2710/16734A61K2039/552A61K2039/5252
Inventor 郑良益漆世华石宝兰朱薇李晶梅舒银辉谢红玲王治江徐松李婷婷
Owner WUHAN CHOPPER BIOLOGY
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