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Porcine pseudorabies virus, and vaccine composition and applications thereof

A technology of porcine pseudorabies virus and vaccine composition, which is applied in the direction of antiviral agents, viruses/phages, microorganisms, etc., can solve the problems of immune effect and unsatisfactory antibody duration, and achieve good immunogenicity, high antibody level, and sustained long term effect

Active Publication Date: 2013-12-11
JIANGSU ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the immune effect and antibody duration of the porcine pseudorabies inactivated vaccine that exist in the prior art are not ideal

Method used

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  • Porcine pseudorabies virus, and vaccine composition and applications thereof
  • Porcine pseudorabies virus, and vaccine composition and applications thereof
  • Porcine pseudorabies virus, and vaccine composition and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 The acquisition and characteristic determination of porcine pseudorabies virus

[0034] 1. Virus Isolation

[0035] The spleen, lung, kidney, and lymphoid tissues of affected piglets from a pig farm in Nanjing, Jiangsu, my country were collected as toxic materials. Put the toxic material in a mortar for aseptic operation first, then add 5ml of sterilized physiological saline, and grind it repeatedly; add 200U of penicillin and 200U of streptomycin to each ml of the treated toxic material, mix well, and mix it Put into -20 ℃ refrigerator and freeze and thaw repeatedly 3 times, freeze and thaw quickly each time to release the virus from the broken cells; centrifuge at 12000 r / min for 10min, take the supernatant and inoculate it into BHK21 cells (purchased from ATCC, That is, the American Type Culture Collection) culture. After 2 days of culture, with normal BHK21 cells ( figure 2 ) compared with that of BHK21 cells inoculated with toxic materials showed obvi...

Embodiment 2

[0056] Example 2 Preparation of vaccine composition containing inactivated porcine pseudorabies virus PRV-JS strain

[0057] 1. BHK21 cell seed propagation and expansion culture

[0058] Take out the frozen BHK21 cell tube from the liquid nitrogen tank, put it in a 37°C water bath to thaw quickly, transfer the cells into a centrifuge tube containing 10ml of DMEM medium, and centrifuge at 1000rpm for 5 minutes. Discard the supernatant, suspend the cells with the growth medium, then add to the cell culture flask, at 37°C, 5% CO 2 cultivated under conditions. When the cell coverage reached 100%, the cells were digested with 0.1% trypsin-EDTA solution. Subculture at a volume ratio of 1:3. The growth medium is DMEM solution containing 10% FBS (newborn bovine serum).

[0059] 2. Virus culture

[0060] The BHK21 cell culture with a cell growth coverage rate of 80% was removed from the growth medium. Inoculate PRV-JS strain virus solution according to the multiplicity of infecti...

Embodiment 3

[0070] Example 3 Safety inspection of porcine pseudorabies inactivated vaccine

[0071] Check the safety of the porcine pseudorabies inactivated vaccine prepared according to the method of Example 2.

[0072] Each batch of vaccines uses 30-35 day-old healthy piglets and healthy pregnant sows negative for pseudorabies antibodies, and each head is intramuscularly injected with 2 parts (4ml) of porcine pseudorabies inactivated vaccine, and another set of the same type that is not vaccinated Pigs were used as control group.

[0073] Observed for 14 days after inoculation, all experimental pigs observed indicators including body temperature, energy, appetite and other local and systemic reactions, the results are shown in Table 2. It can be seen from Table 2 that all test pigs showed no abnormality in spirit, body temperature, appetite, etc. The vaccinated piglets had no abnormal reaction caused by vaccination; the vaccinated pregnant sows had normal pregnancy and no abortion occ...

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Abstract

The invention provides a p porcine pseudorabies virus and vaccine composition and applications thereof, belonging to the field of biotechnology. The microbial preservation number of the porcine pseudorabies virus PRV-JS strain is CGMCCNO.6604. The invention further provides a vaccine composition which comprises an inactivated porcine pseudorabies virus PRV-JS strain and adjuvant acceptable on veteriary pharmacy. The vaccine composition further comprises a carrier acceptable on the veterinary pharmacy. The porcine pseudorabies virus PRV-JS strain is screened from porcine pseudorabies prevalent strains separated from all pig farms and has good immunogenicity, and can be used as inactivated vaccine production virus seeds or virus seeds for testing. After being immunized by the vaccine composition, pigs have higher produced antibody level and the lasting period is long. The vaccine composition prepared by adopting the porcine pseudorabies virus PRV-JS strain can be used for preventing the sow abortionbreeding difficulty and mortality syndromeboar infertility caused by the porcine pseudorabies virus, boar infertility and pseudorabies of other pigs.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a strain of porcine pseudorabies virus, a vaccine composition and applications thereof. Background technique [0002] Porcine pseudorabies virus (Pseudorabies virus) belongs to herpesviridae herpesvirus subfamily porcine herpesvirus type Ⅰ, which can cause acute infectious porcine pseudorabies (Aujeszky's disease). Pseudorabies is widespread in the world, which is extremely harmful to the global pig industry and causes huge economic losses. After pseudorabies occurs in pigs, the clinical symptoms depend on the age of infected pigs, the virulence of virus strains, the dose and route of infection. Adult pigs are generally recessively infected, and pregnant sows can cause syndromes such as abortion, stillbirth, mummification, and infertility in breeding pigs. The morbidity and mortality of piglets within 15 days of age can reach 100%, the incidence of weaned piglets is 40%...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/00A61K39/245A61P31/22C12R1/93
Inventor 唐波张道华张雪花侯继波
Owner JIANGSU ACAD OF AGRI SCI
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