Low virulent strain of porcine reproductive and respiratory syndrome virus, immunogenicity immunogenicity material and vaccine

A technology for respiratory syndrome and pig reproduction. It is applied in the directions of virus antigen components, antiviral agents, viruses/phages, etc. It can solve the problems that the immune efficacy needs to be improved, the pigs cannot be directly vaccinated, and the protection effect is different.

Active Publication Date: 2010-06-16
CHINA ANIMAL DISEASE CONTROL CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] 1. Non-highly pathogenic PRRS virus is generally used as vaccine strain at home and abroad, and the prepared immunogenic substances and vaccines can only resist the attack of highly pathogenic PRRS virus to a certain extent. However, there are differences in the protective effect of products from different manufacturers, and the immune effect is not reliable. It has not been approved by the state to prevent and control the current epidemic of highly pathogenic PRRS.
[0010] 2. The applicant previously prepared immunogenic substances from the highly pathogenic porcine blue-ear disease virus NVDC-JXA1 strain and formulated them into inactivated vaccines, which were designated by the state for the emergency prevention and control of highly pathogenic porcine blue-ear disease in the country. Remarkable results have been achieved, and significant contributions have been made to control the spread of the epidemic, but the disadvantages are: (1) virulent strains can cause disease, and can only be used to produce inactivated immunogenic substances and inactivated vaccines, and cannot be directly administered to pigs (2) Prepare inactivated immunogenic substances and inactivated vaccines with virulent strains, the ability to stimulate the body to produce cellular immune responses is worse than that of live viruses, and the immune efficacy needs to be improved; (3) The production of immunogenic substances and vaccines with virulent strains requires a large amount of virulent cultivation, so the production environment needs to be strictly controlled to prevent the spread of virulent viruses; (4) the cost of preparing inactivated vaccines is much higher than that of attenuated live vaccines

Method used

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  • Low virulent strain of porcine reproductive and respiratory syndrome virus, immunogenicity immunogenicity material and vaccine
  • Low virulent strain of porcine reproductive and respiratory syndrome virus, immunogenicity immunogenicity material and vaccine
  • Low virulent strain of porcine reproductive and respiratory syndrome virus, immunogenicity immunogenicity material and vaccine

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Virus Subculture and Identification Test

[0051] Porcine reproductive and respiratory syndrome virus NVDC JXA1-R attenuated strain (virus content 10 5.0 TCID 50 / ml above), Marc-145 cell line, all provided by the Veterinary Diagnosis Office of China Animal Disease Prevention and Control Center.

[0052] Take well-grown MARC-145 cells, digest them with trypsin, inoculate them in cell flasks, pour out the growth medium after growing into a single layer, replace them with cell maintenance medium containing 5% virus seeds, and culture them at 37°C for 2-3 days. On the first day, when 70% of the cells had lesions, they were harvested, frozen and thawed twice at -40°C, centrifuged, subpackaged, and then identified.

[0053] Using the RT-PCR method, the genome of the attenuated NVDC JXA1-R strain was amplified in segments (different generations of cultures were amplified separately). The obtained gene amplification product is recovered, purified, connected to a sequencing ...

Embodiment 2

[0073] Pathogenicity determination and immunogenicity test of different generations of viruses

[0074] Experimental animals: A total of 30 dual or triple hybrid pigs aged 4 to 6 weeks were taken, and both the antigens detected by PRRS virus RT-PCR kit and the antibodies detected by ELISA were negative. They were randomly divided into 5 pigs / group and raised separately.

[0075] Pathogenicity determination: Inoculate a group of pigs (5 pigs) with 25th, 49th, 60th, and 80th generation PRRS virus JXA1-R strain cultures, and inject 2 mL intramuscularly into each head. The remaining 5 pigs (unvaccinated) served as the control group. The clinical manifestations of pigs were observed and recorded daily until 28 days after inoculation.

[0076] Immunogenicity test: on the 28th day after immunization, a total of 20 pigs inoculated with the PRRS virus JXA1-R strain, and 5 pigs (not inoculated) in the control group were all tested with virulent ( PRRS virus NVDC-JXA1 strain 5th gener...

Embodiment 3

[0083] Pathogenicity determination and immunogenicity test of mixed viruses of different generations

[0084] Experimental animals: A total of 40 dual or triple hybrid pigs aged 4 to 6 weeks were taken, and both the antigens detected by the PRRS virus RT-PCR kit and the antibodies detected by ELISA were negative. They were randomly divided into 5 pigs / group and raised separately.

[0085] Pathogenicity determination: PRRS with 25+49 generations, 25+65 generations, 25+80 generations, 49+80 generations, 49+60 generations, 60+80 generations, 25+49+60+80 generations mixed in equal proportions The culture of the virus JXA1-R strain was inoculated into a group of pigs (5 pigs), and each head was injected intramuscularly with 2 mL. The remaining 5 pigs (unvaccinated) served as the control group. The clinical manifestations of pigs were observed and recorded daily until 28 days after inoculation.

[0086] Immunogenicity test: on the 28th day after immunization, a total of 35 pigs i...

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Abstract

The application relates to a domesticated porcine reproductive and respiratory syndrome virus low virulent strain (JXA1-R strain), immunogenicity substance containing the viral strain, and a porcine reproductive and respiratory syndrome virus low virulent live vaccine developed through making use of the strain. The live vaccine is used to prevent the porcine reproductive and respiratory syndrome,in particular the highly pathogenic blue-ear porcine disease.

Description

technical field [0001] The invention relates to the prevention and control of animal diseases, in particular to an artificial attenuated virus strain of porcine reproductive and respiratory syndrome virus and immunogenic substances and vaccines containing the virus strain. Background technique [0002] Porcine reproductive and respiratory syndrome (also known as porcine blue ear disease) is caused by porcine reproductive and respiratory syndrome virus (porcinereproductive and respiratory syndrome virus, PRRSV), with fever, abortion, and increased mortality of piglets before and after weaning. Diseases with clinical features such as breathing disorder in pigs of different ages. The disease was first discovered in North America in 1987, and almost at the same time, the disease was also reported in Europe. At present, the disease is one of the important diseases seriously affecting the development of the pig industry in the world. [0003] Vaccination is the most effective wa...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/01A61K39/12A61P31/14C12R1/93
Inventor 田克恭
Owner CHINA ANIMAL DISEASE CONTROL CENT
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