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Bivalent inactivated vaccine for porcine reproductive and respiratory syndrome and preparation method thereof

A technology for bivalent inactivated vaccines and respiratory syndrome, applied in multivalent vaccines, biochemical equipment and methods, vaccines, etc., can solve problems such as insufficient immune protection, weak cross-protection, and infection in pigs, and increase the number of pigs Field economic benefits, easy operation, and strong immune protection effect

Pending Publication Date: 2021-10-22
HANGZHOU JIANLIANG VETERINARY BIOLOGICAL PREPARATIONS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The genome similarity between NAD30-like and the currently popular PRRSV vaccine strains is only 85.5-89.5%, resulting in weak cross-protection of commercial vaccines against the current popular NAD30-like vaccine strains, and pig farms immunized with classic PRRSV vaccines and HP-PRRSV vaccines had similar occurrences. The phenomenon of NAD30 strain infection, to some extent, indicates that the existing commercial vaccines cannot produce sufficient immune protection for pigs

Method used

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  • Bivalent inactivated vaccine for porcine reproductive and respiratory syndrome and preparation method thereof
  • Bivalent inactivated vaccine for porcine reproductive and respiratory syndrome and preparation method thereof
  • Bivalent inactivated vaccine for porcine reproductive and respiratory syndrome and preparation method thereof

Examples

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Effect test

Embodiment 1

[0024] Embodiment 1: Isolation and identification of PRRSV epidemic strains

[0025] (1) Viruses, cells and vaccines

[0026] PRRSV isolation epidemic strains ZJ / H and ZJ / NB, PBMC cells, Marc145 cells, porcine peripheral blood mononuclear cell isolation kit, highly pathogenic porcine reproductive and respiratory syndrome live vaccine (JXA1-R strain), porcine Inactivated reproductive and respiratory syndrome vaccine (CH-1a strain).

[0027] Collection and processing of disease materials. Take the fresh lungs and lymph nodes of clinically suspected PRRS-affected pigs, add cold PBS to grind, centrifuge the grinding solution at 3000r / min for 10min, take the supernatant, freeze and thaw twice, and filter with a 0.22μm bacterial filter to obtain the suspected PRRSV virus stock solution.

[0028] Preparation of PBMC cells. First, use an anticoagulant-added anticoagulant tube to slowly extract an equal amount of anticoagulant blood (whole blood: anticoagulant = 6:1), gently invert ...

Embodiment 2

[0044] Embodiment 2: Preparation of Porcine Reproductive and Respiratory Syndrome Bivalent Inactivated Vaccine

[0045] Preparation of virus fluid for vaccines. Take the Marc145 cells that grow into a good monolayer, discard the old culture medium, and inoculate porcine reproductive and respiratory syndrome ZJ / H strain and ZJ / NB strain virus seeds respectively according to the ratio of 5% (V / V), 37 ° C, 5% CO 2 After 1 hour of adsorption in the incubator, DMEM maintenance solution containing 2% fetal bovine serum was added to continue the cultivation. Harvest the virus liquid when 80% of the cells pile up and fall off, freeze and thaw twice, and store at -70°C.

[0046] viral median infective dose (TCID) 50 ) determination. Do 10-fold serial dilution of the above-prepared virus liquid with serum-free medium, and take 10 -4 、10 -5 、10 -6 3 dilutions were inoculated in a 96-well plate of Marc145 cells, 6 wells for each dilution, 0.1ml / well, at 37°C, 5% CO 2 Observation i...

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Abstract

The invention discloses a bivalent inactivated vaccine for porcine reproductive and respiratory syndrome and a preparation method thereof. The bivalent inactivated vaccine for the porcine reproductive and respiratory syndrome comprises the following two inactivated viruses: a porcine reproductive and respiratory syndrome virus with the strain number of PRRSV-ZJ / H strain and the preservation number of CCTCC NO: V202130, and a porcine reproductive and respiratory syndrome virus with the strain number of PRRSV-ZJ / NB strain and the preservation number of CCTCC NO: V202140. The bivalent inactivated vaccine prepared by the invention can effectively prevent and control two major prevalent PRRS virus types in the current field, and a needle of the PRRS inactivated vaccine is immunized in a commercial pig field which is naturally infected or is immunized by using a PRRS attenuated live vaccine, so that the virus specificity neutralizing antibody can be obviously improved, the virus load in the infected pig body can be reduced, the PRRS epidemic situation can be quickly and effectively controlled, the daily gain of pigs can be obviously improved, and the economic benefits of pig farms are increased.

Description

technical field [0001] The invention relates to the field of biological products, in particular to a porcine reproductive and respiratory syndrome bivalent inactivated vaccine and a preparation method thereof. Background technique [0002] Porcine reproductive and respiratory syndrome (PRRS), also known as "pig blue ear disease", is an acute, highly contagious viral infectious disease characterized by reproductive disorders and respiratory symptoms in pigs . The disease can cause miscarriages, premature births, stillbirths, weak piglets in late gestation sows, and dyspnea, high mortality, and slow growth in piglets and growing-finishing pigs. Pigs of all ages are affected. Necropsy showed interstitial pneumonia and enlarged lymph nodes, among which the virus content in tonsils, lungs and lymph nodes was high. [0003] The pathogen of PRRS (Porcine reproductive and respiratory syndrome virus, PRRSV) is a positive-sense small RNA virus belonging to the Arteriviridae family. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/295A61K39/12A61P31/14C12N7/00C12N7/02
CPCA61K39/12A61P31/14C12N7/00A61K2039/552A61K2039/5252A61K2039/70C12N2770/10034C12N2770/10051
Inventor 李少丽王家福尹忠良朱国坡邵攀峰胡晓凤
Owner HANGZHOU JIANLIANG VETERINARY BIOLOGICAL PREPARATIONS CO LTD
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