Porcine Seneca Valley Virus high titer positive serum and preparation method thereof

A pig Seneca, positive serum technology, applied in biochemical equipment and methods, viruses, viruses/phages, etc. Nika Valley virus hyperimmune serum and other issues, to achieve the effect of reducing the risk of foreign viruses, providing a large amount of serum, and reducing side effects in immunized animals

Active Publication Date: 2022-07-29
CHINA ANIMAL HUSBANDRY IND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there is no commercial porcine Seneca Valley virus hyperimmune serum, and there is no relevant report on the preparation method of porcine Seneca Valley virus-specific high-titer positive serum

Method used

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  • Porcine Seneca Valley Virus high titer positive serum and preparation method thereof
  • Porcine Seneca Valley Virus high titer positive serum and preparation method thereof
  • Porcine Seneca Valley Virus high titer positive serum and preparation method thereof

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preparation example Construction

[0032] The present invention provides a preparation method of porcine Seneca Valley virus specificity and high titer positive serum. Guinea pigs are used as immunized animals, virus liquid of suspension culture cells is used as porcine Seneca Valley virus antigen, and porcine Seneca Valley virus antigen is used. The immune response of guinea pigs was stimulated by the synergistic effect of liquid and inactivated vaccine to obtain positive serum with high neutralization titer.

[0033] The test animals used in the present invention are 350-450 g guinea pigs, and the serum neutralizing antibody titer of the guinea pigs to the swine Seneca Valley virus is not higher than 1:4, and the porcine circovirus type 2 antibody and the porcine blue ear virus antibody are detected. Negative, swine Seneca Valley virus, foot-and-mouth disease virus, African swine fever virus antigen test was negative.

[0034] The present invention provides a preparation method of porcine Seneca Valley virus ...

Embodiment 1

[0070] Example 1 Preparation method of porcine Seneca Valley virus specific high titer positive serum

[0071] 1. Preparation of Porcine Seneca Valley Virus Solution

[0072] After the cells were recovered, they were cultured in suspension in a shaker at 37±0.5°C with a cell density of 4×10 6 When the cells / mL is above, the culture is scaled up step by step. Cell density up to 4×10 6 cells / mL, and the cell viability is above 90%, inoculate the cultured suspension cells with 0.1% of the cell fluid volume of the production virus, and set the culture parameters (pH value, DO value, stirring revolution) at 37 Culture at ±0.5°C, take samples for 24 hours, observe under microscope, when the cell viability is less than 20%, harvest the virus culture, freeze and thaw once (freeze-thaw purpose: place the harvested virus liquid at -18°C for overnight, Then thaw at room temperature to completely release the virus from the cells), and store it below -40°C.

[0073] Preparation of Porc...

Embodiment 2

[0083] Embodiment 2 The preparation method of porcine Seneca Valley virus specific high titer positive serum

[0084] 1. Preparation of Porcine Seneca Valley Virus Antigen Solution

[0085] After the cells were recovered, they were cultured in suspension in a shaker at 37±0.5°C with a cell density of 4×10 6 When the cells / mL is above, the culture is scaled up step by step (pH7.2±0.1, dissolved oxygen DO 30%~60%, stirring speed 60~150rpm). Cell density up to 4×10 6cells / mL, and the cell viability is above 90%, inoculate the cultured suspension cells with 0.5% of the cell fluid volume, and set the culture parameters (pH7.2±0.1, dissolved oxygen DO 40%~ 60%, stirring speed 80-110rpm) at 37±0.5°C, take samples for 24 hours, observe under the microscope, when the cell viability is less than 20%, harvest the virus culture, freeze and thaw once (freeze-thaw purpose: the harvested The virus solution was kept at -18°C overnight, and then thawed at room temperature to completely rele...

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Abstract

The invention provides a porcine Seneca Valley virus high-titer positive serum and a preparation method thereof. The method includes: obtaining a positive serum with a high neutralization titer through the synergistic action of a pig Seneca Valley virus antigen liquid and an inactivated vaccine to stimulate an immune response produced by an experimental animal (such as a guinea pig), and the prepared Seneca Valley specificity The titer of positive serum is as high as 1:1024 or more, which can be used for the specificity test, differential test, exogenous virus test and diagnostic detection of Seneca Valley virus vaccine, etc. new ideas.

Description

technical field [0001] The invention relates to the field of biological products, in particular to a porcine Seneca Valley virus high-titer positive serum and a preparation method thereof. Background technique [0002] Seneca valley virus (SVV), also known as Seneca virus A (SVA), is the only member of the picornavirus family Senecavirus genus. SVV infection can cause vesicular disease in pigs. After the virus is infected, vesicle-like lesions appear on the snout and hoof crown of pigs, with occasional diarrhea symptoms, and the condition is acute. The mortality rate of newborn piglets is 30% to 70%. SVV is an emerging animal pathogen with unknown risks, and the development of its diagnostic products and serum products has become a top priority. At present, there is no commercialized porcine Seneca Valley virus hyperimmune serum, and there is no relevant report on the preparation method of porcine Seneca Valley virus-specific high-titer positive serum. SUMMARY OF THE INVE...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/06C07K16/10C12N7/00C12R1/93
CPCC07K16/06C07K16/1009C12N7/00C07K2317/35C12N2770/32051
Inventor 吴珊珊王贵华张贺楠杨君敬韩伟
Owner CHINA ANIMAL HUSBANDRY IND
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