A kind of porcine pseudorabies virus strain, vaccine composition and its preparation method and application

A technology of porcine pseudorabies and vaccine composition, applied in the field of animal virology, capable of solving problems such as respiratory tract and/or neurological symptoms

Active Publication Date: 2018-08-31
PU LIKE BIO ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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 pseudorabies after injection of commercial vaccines in piglets, such as prolonged body temperature rise, depression, loss of appetite, respiratory tract and / or neurological symptoms

Method used

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  • A kind of porcine pseudorabies virus strain, vaccine composition and its preparation method and application
  • A kind of porcine pseudorabies virus strain, vaccine composition and its preparation method and application
  • A kind of porcine pseudorabies virus strain, vaccine composition and its preparation method and application

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. Primer Premier 5.0 was used to design primer sequences for amplifying the gD gene, respectively.

[0056] Using the extracted genomic DNA as a template, prepare the PCR amplification system as follows: template DNA 100 μg, PrimerSTARHS DNA Polymerase (2.5 U / μl) 0.5 μl, 2×PrimerSTARGC Buffer 25 μl, upstream and downstream primers 1 μl (10 pmol / μl), dNTP Mix (2.5 mM 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 analyzed b...

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, which comprises porcine pseudorabies virus antigen in an immune amount and an adjuvant acceptable in veterinary medicine. The vaccine composition can induce an effective immune response in pigs and dogs, and piglets immunized with the vaccine composition have shorter body temperature rise time, basically normal appetite, and basically no clinical symptoms, showing 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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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/00C12N15/38A61K39/245A61P31/22C12R1/93
Inventor 李向东谭菲菲肖燕张许科孙进忠田克恭
Owner PU LIKE BIO ENG
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