Swine pseudorabies virus strain, vaccine composition, preparation method and application thereof

A technology for porcine pseudorabies and vaccine composition, which is applied in the field of animal virology and can solve the problems of respiratory and/or neurological symptoms and the like

Active Publication Date: 2015-02-04
PU LIKE BIO ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] There is only one serotype of pig PRV, and it is generally believed that the cross-protection of the strain is very strong, but there are still typical porcine pseudora

Method used

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  • Swine pseudorabies virus strain, vaccine composition, preparation method and application thereof
  • Swine pseudorabies virus strain, vaccine composition, preparation method and application thereof
  • Swine pseudorabies virus strain, vaccine composition, preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Embodiment 1, collection and isolation of virus

[0052] Isolate the sample from the suspected porcine pseudorabies infection sample from Henan, aseptically collect pig brain tissue, add MEM culture solution at 1:10 (volume ratio), grind, prepare tissue suspension, after repeated freezing and thawing for 3 times, 2000r / min centrifuged for 15 min, collected the supernatant, and then filtered through a 0.2 μm membrane filter, inoculated on PK-15 cells and incubated at 37°C for 1 hour, replaced with MEM medium containing 2% calf serum, and incubated at 37°C for 5 days. Harvest the poisonous culture medium, freeze and thaw twice, collect the poison, and add MEM culture medium containing 2% calf serum. Porcine pseudorabies virus PCR detection kit (Beijing Century Yuanheng Animal Epidemic Prevention Technology Co., Ltd.) was used to detect porcine pseudorabies virus, and the result was positive; the isolated virus was detected by using PCR kit to detect exogenous virus (porc...

Embodiment 2

[0054] Embodiment 2, the genetic characteristic of isolated virus

[0055] The genetic characteristics of the virus isolated in Example 1 were determined by genetic analysis. The genomic DNA of porcine pseudorabies virus isolated on PK15 cells was used as a template, and the primers shown in Table 1 were used for PCR. The primer sequences for amplifying the gD gene were designed using Primer Premier 5.0, respectively.

[0056] Using the extracted genomic DNA as a template, the PCR amplification system was prepared as follows: template DNA 100 μg, PrimerSTAR HS DNA Polymerase (2.5U / μl) 0.5 μl, 2×PrimerSTAR GC Buffer 25 μl, upstream and downstream primers 1 μl (10 pmol / μl), dNTP Mix ( 2.5mM each) 4 μl, and make up the volume to 50 μl with distilled water. A two-step PCR reaction was performed: denaturation at 98°C for 10sec, followed by annealing and extension at 68°C for 1min15sec, a total of 30 cycles. The PCR reaction was terminated at 4°C. The resulting PCR products were...

Embodiment 3

[0059] Embodiment 3, the pathogenicity test of virus

[0060] 3.1 Pathogenicity of piglets of different ages

[0061] Six 34- to 35-day-old swine pseudorabies antibody-negative piglets were randomly divided into 2 groups, 5 pigs / group (test group) and 2 pigs / group (control group), and were inoculated with porcine pseudorabies virus HN1202 strain (challenge dose 2×10 8.0 TCID 50 per head), the control group was inoculated with DMEM medium; after the virus inoculation, the body temperature of the piglets was measured every day, and the clinical symptoms and death were observed. The specific results are shown in Table 2.

[0062] Table 2 Pathogenicity of swine pseudorabies virulent HN1202 strain to piglets of different ages

[0063]

[0064] The results showed that porcine pseudorabies virus HN1202 strain inoculated 35-day-old piglets could cause disease and 3 / 5 piglets died.

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Abstract

The invention provides a vaccine composition. The vaccine composition comprises an immune amount of a swine pseudorabies virus antigen and a veterinary medicine-acceptable adjuvant. The vaccine composition can induce effective immune reactions in the swine and dog. A piglet immunized by the vaccine composition has short body temperature-raising time and substantially normal appetite, without clinical symptoms. The vaccine composition presents good immune protection.

Description

technical field [0001] The invention relates to a vaccine composition and belongs to the field of animal virology. Background technique [0002] Pseudorabies, also known as Aujeszky's disease, is a disease of pigs, cattle, sheep and other domestic animals, poultry and wild animals caused by Suid herpesvirus 1 strain in the α subfamily of the Herpesviridae (Herpesviridae) family. An acute infectious disease characterized by fever, severe itching (except pigs) and encephalomyelitis. Pseudorabies in pigs is widespread in our country and is seriously harmful. It is one of the main diseases restricting the production of large-scale pig farms. It can cause abortion in pregnant sows, stillbirth or mummified fetuses and piglets with neurological symptoms, paralysis, and high mortality. PRV has strong pantropicity, neurotropicity, and latent infection characteristics. It can be latently infected in the peripheral nervous system for a long time. When the latent virus is activated an...

Claims

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Application Information

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IPC IPC(8): C12N7/00C12N15/38A61K39/245A61P31/22C12R1/93
Inventor 张许科孙进忠田克恭
Owner PU LIKE BIO ENG
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