Porcine pseudorabies virus with six-gene deletion, porcine pseudorabies vaccine and preparation method

A porcine pseudorabies virus and porcine pseudorabies technology, which is applied in the fields of porcine pseudorabies virus, porcine pseudorabies vaccine and preparation, can solve the problems of unsatisfactory immunization effect of porcine pseudorabies vaccine, and achieve the effect of good immune effect

Active Publication Date: 2019-04-12
SICHUAN HUASHEN ANIMAL BIOLOGICAL PRODS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the immune effect of the existing porcine pseudorabies vaccine is not ideal

Method used

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  • Porcine pseudorabies virus with six-gene deletion, porcine pseudorabies vaccine and preparation method
  • Porcine pseudorabies virus with six-gene deletion, porcine pseudorabies vaccine and preparation method
  • Porcine pseudorabies virus with six-gene deletion, porcine pseudorabies vaccine and preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Artificially Modified Pseudorabies Virus

[0043] 1. Virus strain

[0044] The PRV-FJ strain was isolated from the Sichuan Provincial Key Laboratory of Livestock and Poultry Infectious Diseases from the pig disease materials of Fujian pig pseudorabies in 2015; the diseased pigs mainly showed symptoms such as elevated body temperature, poor appetite, and sneezing. PRV-FJ strain with 10 7 TCID 50 The dose per head infected 28-day-old weaned piglets without neurological symptoms, and all died 7 days after the challenge.

[0045] ST cells and 293T cells were cultured in DMEM medium containing 10% fetal bovine serum (Gibco).

[0046] 2. Construction of recombinant vector

[0047] Artificially synthesized SEQ ID NO.1, SEQ ID NO.2, and SEQ ID NO.3, then directional cloned SEQ ID NO.1 into the Ase1 restriction site of the pEGFP-C1 vector, and named it pEGFP-gI, SEQ ID NO.2 was directed to the Mlu1 restriction site of pEGFP-gI, and it was named pEGFP-gI28K. SEQ ID NO.3 was d...

Embodiment 2

[0062] PRV-TIE18G Cell Adaptability and Purity Test

[0063] Take the preserved PRV-TIE18G virus, inoculate BHK21 cells by conventional methods, cultivate until CPE appears, and harvest the virus supernatant. The PRV-TIE18G virus was serially passaged to F5. Viral TCID50 assays were performed on BHK21 cells. According to the method recorded in the appendix of "Chinese Veterinary Pharmacopoeia" (2015 edition), different generations of viruses were tested for sterility, mycoplasma and exogenous viruses.

[0064] Table 2 PRV-TIE18G cell adaptability and purity test

[0065] Virus generation

[0066] PRV-TIE18G virus F0 generation virus content is 10 7.6 TCID 50 / ml; continuous passage to the F5 generation, the virus content of the F1 generation to the F5 generation was all within 10 8.0 TCID 50 / ml or more. The results of sterility test and mycoplasma test were negative, and the results of exogenous virus test showed no exogenous virus contamination.

Embodiment 3

[0068] Pathogenicity test of PRV-TIE18G

[0069] 35 PRV antibody-negative piglets aged 1 to 3 days were randomly divided into 6 groups marked as A, B, C, D, E, F, and G groups, and the F1 generation of PRV-TIE18G virus was selected according to the following table (Table 3) Conduct an attack test. Continuous observation for 14 days after the challenge.

[0070] Table 3 Pathogenicity test of PRV-TIE18G and PRV-FJ to newborn piglets

[0071] group

Challenge strain

Virus generation

Attack dose

attack method

Remark

A

PRV-TIE18G

F1

10 8 TICD 50

Intranasal vaccination

B

PRV-TIE18G

F1

10 7 TICD 50

Intranasal vaccination

C

PRV-TIE18G

F1

10 8 TICD 50

intramuscular injection

D

PRV-TIE18G

F1

10 7 TICD 50

intramuscular injection

E

DMEM medium

\

2.0ml

Nasal drops + intramuscular injection

blank control

F

PRV-FJ ...

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Abstract

The invention discloses a porcine pseudorabies virus with six-gene deletion, a porcine pseudorabies vaccine and a preparation method, and relates to the field of animal biological products. The accession number of the porcine pseudorabies virus with six-gene deletion provided by present invention is CGMCC No:16290. The porcine pseudorabies virus with six-gene deletion lacks six genes, the pathogenicity is removed, and the immunogenicity is effectively retained. When in use, the vaccine prepared by the porcine pseudorabies virus can induce a higher dose of interferon in the body, and has a better immune effect in emergency immunization.

Description

technical field [0001] The invention relates to the field of porcine pseudorabies vaccine, in particular to a six-gene-deleted porcine pseudorabies virus, porcine pseudorabies vaccine and a preparation method. Background technique [0002] Porcine pseudorabies (Pseudorabies, PR), also known as Aujeszky's disease (Aujeszky's disease, AD), is caused by pseudorabies virus (Pseudorabies virus, PRV) in pigs, cattle, sheep and other domestic and wild animals with fever , itching (except pigs) and encephalomyelitis as the main symptoms of a highly contagious infectious disease. Pigs are the natural host, main storage host and source of infection of the disease, whether they are newborn piglets, finishing pigs, or adult breeding pigs, all are threatened by PR. The symptoms are: the mortality rate of piglets within 2 weeks of age after infection is very high, up to 100%, diarrhea, vomiting, ataxia, opisthotonos, limb swimming and other neurological symptoms may occur, and finally pa...

Claims

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Application Information

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IPC IPC(8): C12N7/00A61K39/245A61K47/26A61K47/18A61K47/42A61P31/22C12R1/93
CPCA61K39/12A61K47/183A61K47/26A61K47/42A61K2039/522A61K2039/552A61P31/22C12N7/00C12N2710/16721C12N2710/16734
Inventor 周远成林艳代洪波王雪涛廖果邝声耀阴文奇
Owner SICHUAN HUASHEN ANIMAL BIOLOGICAL PRODS
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