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Porcine reproductive and respiratory syndrome virus strain and application thereof

A technology for respiratory syndrome and pig breeding, which is applied in the direction of viruses, antiviral agents, and viral antigen components. It can solve problems such as mixed infection, complex disease, and unsatisfactory prevention effect of PRRSV, so as to improve productivity and improve immunogenicity. sexual effect

Active Publication Date: 2017-07-07
PU LIKE BIO ENG
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

And the preventive effect of existing attenuated live vaccine is not very ideal clinically, although the inactivated vaccine is also developed earlier, it does not have the risk of loose poison and virulence returning to strong, but it is not very effective for the prevention of new PRRSV Ideally, moreover, PRRSV is often co-infected with other pathogens, resulting in the complexity of the clinical condition. Therefore, it is necessary to develop a vaccine composition for the latest popular strains in my country to effectively prevent the prevalence of PRRSV-related diseases in my country

Method used

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  • Porcine reproductive and respiratory syndrome virus strain and application thereof
  • Porcine reproductive and respiratory syndrome virus strain and application thereof
  • Porcine reproductive and respiratory syndrome virus strain and application thereof

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Embodiment approach

[0030] As an embodiment of the present invention, the vaccine composition includes the inactivated whole virus antigen of porcine reproductive and respiratory syndrome virus HNjz15 strain or its culture, and the inactivated whole virus antigen content of the HNjz15 strain or its culture is ≥10 before inactivation 6.0 TCID 50 / ml.

[0031] As a preferred embodiment of the present invention; the inactivated whole virus antigen content of the HNjz15 strain or its culture is 10% before inactivation. 6.0 ~10 8.0 TCID 50 / ml.

[0032] As a more preferred embodiment of the present invention, the inactivated whole virus antigen content of the HNjz15 strain or its culture is 10% before inactivation. 7.0 TCID 50 / ml.

[0033] When porcine reproductive and respiratory syndrome virus is less than 10 6.0 TCID 50 When used in low amounts, the vaccine is not effective in stimulating antibody production. On the other hand, excess amounts may be uneconomical.

[0034] One aspect of ...

Embodiment 1

[0073] The collection and isolation of embodiment 1 virus

[0074]Isolate from samples suspected of porcine reproductive and respiratory syndrome virus infection from Henan, collect porcine tonsil lymphoid tissue aseptically, add DMEM culture medium at 1:10 (volume ratio), grind, prepare tissue suspension, repeat 3 times After freezing and thawing, centrifuge at 12000r / min for 15min, collect the supernatant, filter it through a 0.22μm membrane filter, culture it on PAM cells at 37℃ for 1h, replace it with DMEM culture medium containing 2% calf serum, and culture at 37℃ 5th. Harvest the poisonous culture medium, freeze and thaw twice, collect the poison, and add DMEM culture medium containing 2% calf serum. The porcine reproductive and respiratory syndrome virus PCR detection kit (Beijing Century Yuanheng Animal Epidemic Prevention Technology Co., Ltd.) was used to detect the porcine reproductive and respiratory syndrome virus, and the result was positive; Detection of porcin...

Embodiment 2

[0076] The genetic characteristic of embodiment 2 isolated virus

[0077] The genetic characteristics of the virus isolated in Example 1 were determined by genetic analysis. The genome of porcine reproductive and respiratory syndrome virus isolated on PAM cells was used as a template, reverse-transcribed into cDNA, and PCR was performed using the primers shown in Table 1. Primer Premier 5.0 was used to design primer sequences for amplifying GP5, GP2, and Nsp2 genes, respectively.

[0078] The PCR amplification system is as follows: Template cDNA 1μl, PrimerSTAR HS DNA Polymerase (2.5U / μl) 0.5μl, 5×PrimeSTAR TM Buffer10μl, upstream and downstream primers 1μl (10pmol / μl), dNTP Mix (2.5mM each) 4μl, and ddH 2 O Make up the volume to 50 μl. A two-step PCR reaction was performed: 2 min at 98°C; 10 s at 98°C, 1 min at 55°C, 1 mim / kb at 72°C, 30 cycles; 5 min at 72°C. The resulting PCR products were analyzed by electrophoresis on a 1% agarose gel containing ethidium bromide. PC...

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Abstract

The invention provides a porcine reproductive and respiratory syndrome virus strain and application thereof. The virus strain contains proteinGP5 shown in a sequence table SEQ NO.2 or further contains protein GP2 of which the nucleotide sequence code is shown in a sequence table SEQ NO.3 and protein Nsp2 of which the nucleotide sequence code is shown in asequence table SEQ NO.4. The invention further provides a vaccine composition prepared from the virus strain. The prepared vaccine composition can effectively prevent the affection of new epidemic porcine reproductive and respiratory syndrome viruses (PRRV), stimulate bodies to rapidly generate immunity, and effectively protect swine herds.

Description

technical field [0001] The invention belongs to the field of veterinary vaccines, and in particular relates to porcine reproductive and respiratory syndrome virus strains, vaccine compositions prepared therefrom, preparation methods and applications. Background technique [0002] Porcine reproductive and respiratory syndrome is caused by porcine reproductive and respiratory syndrome virus (porcinereproductive and respiratory syndrome virus, PRRSV), which is characterized by fever, abortion in sows, increased mortality of piglets before and after weaning, respiratory disorders in pigs of different ages, etc. disease with clinical features. The disease was first discovered in North Carolina in the United States (1987), and then successively found in Canada (1998), Germany (1990), and the United Kingdom (1991). In 1992, the International Veterinary Office (OIE) named the disease Porcine Reproductive and Respiratory Syndrome (PRRS). The prevalence of this disease was also repo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00A61K39/12A61K39/155A61K39/02A61K39/102A61K39/295A61P31/20A61P31/04A61P31/16A61P31/14C12R1/93
CPCA61K39/0241A61K39/102A61K39/12A61K2039/5252A61K2039/5254A61K2039/552A61K2039/70C12N7/00C12N2760/16134C12N2760/20134C12N2770/10021C12N2770/10034
Inventor 田克恭李英英孙进忠张许科
Owner PU LIKE BIO ENG
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