Recombinant porcine pseudorabies virus and application thereof and recombinant porcine live pseudorabies vaccine

A porcine pseudorabies virus, pseudorabies virus technology, applied in the direction of veterinary vaccines, vaccines, viruses, etc.

Active Publication Date: 2019-02-15
SICHUAN HUASHEN ANIMAL BIOLOGICAL PRODS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For porcine pseudorabies and porcine epidemic diarrhea, the use of vaccine immunization can significantly reduce the economic loss caused by disease infection, but there is no dual vaccine for porcine pseudorabies and porcine epidemic diarrhea virus on the market

Method used

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  • Recombinant porcine pseudorabies virus and application thereof and recombinant porcine live pseudorabies vaccine
  • Recombinant porcine pseudorabies virus and application thereof and recombinant porcine live pseudorabies vaccine
  • Recombinant porcine pseudorabies virus and application thereof and recombinant porcine live pseudorabies vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Artificially Modified Pseudorabies Virus

[0040] 1. Virus strain

[0041] 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.

[0042] PEDV-CV777 strain and SA014 strain were provided by Sichuan Provincial Key Laboratory of Livestock and Poultry Infectious Diseases.

[0043] ST cells and 293T cells were cultured in DMEM medium containing 10% fetal bovine serum (Gibco).

[0044] 2. Construction of recombinant vector

[0045] Artificially synthesized SEQ ID NO.1, SEQ ID NO.2, SEQ ID NO.3, SEQ ID NO.4, and then directional cloned SEQ ID NO.1 into the Ase I restriction site of th...

Embodiment 2

[0059] Pathogenicity of rPRV-TIE18(RBD)

[0060] Take 20 PRV antibody-negative piglets aged 1-3 days, and randomly divide them into 4 groups, marked as A, B, C, and D groups, and conduct the challenge test according to the table below. Continuous observation for 14 days after the challenge.

[0061] Table 2 Pathogenicity test of rPRV-TIE18(RBD) and PRV-FJ on newborn piglets

[0062] group

Challenge strain

Attack dose

Vaccination method

Remark

A

rPRV-TIE18(RBD)

10 8 TICD 50

Nasal drip

B

rPRV-TIE18(RBD)

10 7 TICD 50

Nasal drip

C

DMEM medium

2.0ml

Nasal drip

blank control

D

PRV-FJ

10 7 TICD 50

Nasal drip

[0063] After the challenge, the temperature of the piglets was observed and measured every day to observe whether there were clinical symptoms and death of pseudorabies. The results are shown in Table 3 below. After rPRV-TIE18 (RBD) was instil...

Embodiment 3

[0067] Preparation and Test of Porcine Pseudorabies Live Vaccine rPRV-TIE18(RBD)

[0068] Inoculate BHK cells with rPRV-TIE18(RBD), and harvest the virus liquid when more than 90% of the cells are damaged. After 2-fold dilution of the harvested virus liquid, it was mixed with gelatin sucrose protectant (containing 2% gelatin and 10% sucrose) in equal volumes, and then divided into 2ml / bottles for freeze-vacuum drying. According to the appendix of "Chinese Veterinary Drug Store", there is no bacterial and mycoplasma pollution, and no exogenous virus pollution. The titer of cultured virus solution is 10 8.0 TCID 50 / ml, the freeze-dried vaccine was restored to the original volume and the virus content was 10 7.25 TCID 50 / ml.

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Abstract

The invention discloses a recombinant porcine pseudorabies virus and application thereof and a recombinant porcine live pseudorabies vaccine, and relates to the field of biological products for animals. The recombinant porcine pseudorabies virus provided by the invention has the collection number of CGMCC No: 16289. The recombinant porcine pseudorabies virus is obtained after artificial transformation, the pathogenicity thereof is removed, pseudorabies virus TK, gI, gE, 11K and 28K genes are deleted, and fusion protein obtained through fusion of porcine epidemic diarrhea virus S truncated protein and porcine CD40L protein can be expressed. After a pig is immunized by the recombinant porcine live pseudorabies vaccine prepared by using the recombinant porcine pseudorabies virus, the pig canacquire the immunity against the porcine epidemic diarrhea virus challenge and the porcine pseudorabies virus challenge. The recombinant porcine live pseudorabies vaccine has a relatively good immuneeffect and has a broad application prospect.

Description

technical field [0001] The invention relates to the field of animal biological products, in particular to a recombinant porcine pseudorabies virus and its application and a live recombinant porcine pseudorabies vaccine. Background technique [0002] Pseudorabies (PR), also known as Aojeszky's disease (Aujeszky's disease, AD), is caused by pseudorabies virus (Pseudorabiesvirus, PRV) in pigs, cattle, sheep and other domestic and wild animals with fever, Itching (except pigs) and encephalomyelitis is a highly contagious infectious disease with main symptoms. Pigs are the natural host, main storage host and source of infection of the disease. PRV can infect pigs of all ages, and the younger the age, the more severe the disease and the higher the mortality rate after infection. The mortality rate of suckling piglets after infection is very high, up to 100%, and neurological symptoms such as diarrhea, vomiting, and swimming of limbs may occur; weaned piglets and fattening pigs ar...

Claims

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Application Information

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IPC IPC(8): C12N7/00A61K39/245A61K47/42A61K47/26A61P31/22
CPCA61K39/12A61K47/26A61K47/42A61K2039/5254A61K2039/5256A61K2039/552A61P31/22C12N7/00C12N2710/16721C12N2710/16734C12N2710/16762
Inventor 周远成代洪波林艳廖果王雪涛邝声耀阴文奇
Owner SICHUAN HUASHEN ANIMAL BIOLOGICAL PRODS
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