Porcine pseudorabies virus, vaccine composition and preparation method and use thereof

a pseudorabies virus and vaccine technology, applied in the field of vaccine composition, can solve the problems of high death rate of piglets, neurological signs, stillborn or mummified fetuses, etc., and achieve the effect of good immune function of porcine pseudorabies

Active Publication Date: 2016-04-28
PU LIKE BIO ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0006]In order to solve the deficiency of the prior art, the present invention aims to provide a porcine pseudorabies virus strain for preparing vaccines, which have been proved by animal testing to provide a good immune function for porcine pseudorabies.

Problems solved by technology

Pseudorabies in swine is found nationwide in China causing severe damages, and is one of the major diseases limiting the large-scale production of pig farms.
Infection can result in abortion, stillborn or mummified fetuses in pregnant sows, and neurological signs, paralysis and a high death rate in piglets.
However, piglets may still suffer from typical porcine pseudorabies after their injection with commercial vaccines, with symptoms such as long-term high fever, depression, loss of appetite, respiratory and / or neurological signs.
There are no vaccines in the prior art capable of solving the pseudorabies caused by variant strains of porcine pseudorabies virus.

Method used

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  • Porcine pseudorabies virus, vaccine composition and preparation method and use thereof

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0059]Collection and Isolation of Viruses

[0060]Porcine brain tissue was collected under aseptic conditions from samples isolated from samples from Henan province suspected of having pseudorabies infection, added in MEM liquid medium in a ratio of 1:10(V:V), and ground to prepare a tissue suspension. After 3 times of repeated freezing-thawing, the tissue suspension was centrifuged at 2000r / min for 15 minutes. The supernatant was then collected, filtered through a 0.2 μm pore filter, subcultured on PK-15 cells and incubated at 37° C. for lh, and then the medium was changed by adding MEM liquid medium supplemented with 2% fetal bovine serum, and incubated at 37° C. for 5 days. PRV was detected by PRV PCR detection kit (Beijing Anheal Laboratories Co., Ltd), and the result was positive; PCR kit was employed to detect the exotic virus contamination (porcine reproductive and respiratory syndrome virus RT-PCR detection kit kit, porcine parvovirus PCR detection kit and classical swine fever...

example 2

[0062]Genetic Characteristics of the Isolated Virus

[0063]Genetic characteristics of the isolated virus in Example 1 were determined by means of gene analysis. Genomic DNA prepared from the pseudorabies virus isolated from PK-15 cells was used as template with primers shown in Table 1 for PCR amplification reactions. The Primer Primer 5.0 was used for designing the primer sequence for amplifying gB, gC and gD gene, respectively.

[0064]The genomic DNA extracted was used as template to prepare the following PCR amplification system: 100 μg template DNA, 0.5 μL PrimerSTAR HS DNA Polymerase (2.5U / μL), 25 μL 2 ×PrimerSTAR GC Buffer, 1 μL of each upstream and downstream primer (10 pmol / μL), 4 μL dNTP Mix (2.5 mM each), adjusted to a final volume of 50 μL with distilled water. Two-step PCR reaction was carried out by an initial denaturation for 10 sec at 98° C. followed by annealing and extension at 68° C. (all the time is calculated by 1 kb / min) and there were 30 cycles in total. The PCR re...

example 3

[0065]Pathogenicity Test of the Virus

[0066]3.1 Pathogenicity of the Virus in Piglets at Different Days of Age

[0067]6 piglets at 34˜35 days of age which were negative for pseudorabies antibodies were randomly divided into 2 groups, one with 4 piglets (experimental group) and the other with 2 piglets (control group), wherein the experimental group was inoculated with PRV HN1201 strain by intranasal instillation (challenge dosage is 2×108.0TCID50 / piglet) and the control group was inoculated with DMEM medium. Meanwhile 4 piglets at 49 days of age were inoculated with third passage of the virulent HN1201 strain after preservation (challenge dose was 2×108.0TCID50 / piglet), and the control is still piglets at 35 days of age. After inoculation of virus, the temperature of piglets was determined daily, and clinical signs and death status were observed. The results are shown in Table 2.

TABLE 2Pathogenicity of PRV HN1201 strain in piglets at different days of ageNum-Inoculation GroupberDaysDos...

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Abstract

Provided in the embodiments of the present invention is a vaccine composition including an immune amount of attenuated live vaccine, inactivated vaccine, subunit vaccine, synthetic vaccine, or genetically engineered vaccine of the porcine pseudorabies virus strain. The vaccine composition can effectively induce antibody generation, and prevent infections of virulent strains of the porcine pseudorabies virus, and provides effective protection for pigs.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS[0001]This application is a continuation of U.S. patent application Ser. No. 14 / 779,132, filed Sep. 22, 2015 which claims priority to International Application No. PCT / CN2014 / 076691, filed Apr. 30, 2014, which claims priority to Chinese Application No. 201310216881.6, filed May 31, 2013 and Chinese Application No. 201310428605.6, filed Sep. 18, 2013, the entire contents of which are hereby incorporated by reference.FIELD OF THE INVENTION[0002]This invention relates to a vaccine composition, belonging to the field of animal virology.BACKGROUND[0003]Pseudorabies, also called Aujeszky's disease, is an acute infectious disease caused by Suid herpesvirus 1 (SuHV1) belonging to the Alphaherpesvirinae subfamily for many kinds of livestock such as swine, cattle and sheep, as well as poultry and wild animals, with the main symptoms of fever, intense itching (except swine) and encephalomyelitis. Pseudorabies in swine is found nationwide in China causing ...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61K39/245C12N7/00C07K14/005
CPCA61K39/245C07K14/005C12N7/00A61K2039/552C12N2710/16734A61K2039/5252C12N2710/16771C12N2710/16722C07H21/04C12N7/04A61K39/12A61P31/22C12N2710/16721C12N2710/16761A61K39/00
Inventor ZHANG, XUKESUN, JINZHONGWU, RUITAN, FEIFEIBAI, CHAOYONGTIAN, KEGONG
Owner PU LIKE BIO ENG
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