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Seneca virus isolation strain, Seneca virus disease inactivated vaccine and preparation method of Seneca virus disease inactivated vaccine

An inactivated vaccine, virus disease technology, applied in the direction of virus antigen components, biochemical equipment and methods, viruses, etc., can solve the problems of no particularly effective means of Seneca virus prevention and control, and the trouble of pig farming.

Pending Publication Date: 2020-03-27
哈药集团生物疫苗有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Pigs of all ages can be infected by the virus, causing huge problems for the swine industry
[0003] At present, there is no particularly effective method for the prevention and control of Seneca virus, and there is no commercial vaccine on the market, so there is an urgent need for a vaccine that can be used to prevent and treat Seneca virus disease in swine

Method used

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  • Seneca virus isolation strain, Seneca virus disease inactivated vaccine and preparation method of Seneca virus disease inactivated vaccine
  • Seneca virus isolation strain, Seneca virus disease inactivated vaccine and preparation method of Seneca virus disease inactivated vaccine
  • Seneca virus isolation strain, Seneca virus disease inactivated vaccine and preparation method of Seneca virus disease inactivated vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1 Isolation and Identification of Seneca Virus CH-HLJ Strain

[0024] 1 test material

[0025] The disease material comes from a pig farm suspected to be infected with Seneca virus in Heilongjiang Province; BHK-21 cells are preserved by Harbin Pharmaceutical Group Bio-vaccine Co., Ltd.

[0026] 2 test method

[0027] 2.1 Collection and processing of disease materials

[0028] Blister fluid was collected from pigs suspected of being infected with SVA. After repeated freezing and thawing, 1% penicillin-streptomycin was added and distributed into 1.5mL sterile EP tubes. Store at ℃ for later use.

[0029] 2.2 Virus detection

[0030] Total RNA was extracted with a commercial DNA / RNA extraction kit, reverse-transcribed to obtain a cDNA template, PCR amplification was performed using a pair of specific primers for the SVA gene, and PCR products were detected by 1% agarose gel electrophoresis.

[0031] Table 1 identifies the primers of the SVA gene

[0032]

...

Embodiment 2

[0051] The preparation of embodiment 2 Seneca virus CH-HLJ strain inactivated vaccines

[0052] 1 virus multiplication

[0053] According to 5‰ of the total amount of the culture medium, inoculate the BHK-21 cells with a single layer of virus seed, and place it at 37°C in 5% CO 2 Culture in the incubator, after adsorption for 1 hour, add DMEM cell culture medium with 2% serum content, continue to store at 37°C in 5% CO 2cultured in an incubator. After inoculation, the cells are cultured for 40-48 hours, and when the cytopathy reaches more than 80%, the cells and the cell culture are harvested.

[0054] 2 semi-finished product inspection

[0055] 2.1 Sterility test

[0056] Tested according to the appendix of the current "Chinese Veterinary Pharmacopoeia", the virus content of the virus liquid used for making seedlings is 9.5TCID 50 / ml, the sterility test is qualified.

[0057] 2.2 Determination of virus content

[0058] The BHK-21 cells in the logarithmic growth phase ...

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Abstract

The invention discloses a Seneca virus isolation strain, Seneca virus disease inactivated vaccine and a preparation method of the Seneca virus disease inactivated vaccine. A swine Seneca virus CH-HLJstrain is separated from a pig farm with symptoms of suspected Seneca virus infections in the Heilongjiang Province, the complete genome sequence of the strain is shown in SEQ ID No.1 in the description, and the microbial preservation number is CGMCC NO.18851. The invention further provides a method for preparing the Seneca virus disease inactivated vaccine. The method comprises the following steps: (1) performing viral multiplication and inactivation; (2) heating a vaccine adjuvant so as to obtain an oil phase; (3) heating a virus liquid so as to obtain a water phase; and (5) mixing and emulsifying the oil phase with the water phase into a double-phase oil emulsion. Swine immunological experiment results show that the Seneca virus disease inactivated vaccine prepared by the invention is capable of stimulating a body to generate an antibody of a high level. Immunoprotection efficacity evaluation results of the inactivated vaccine show that the inactivated vaccine prepared by the invention is capable of providing good protection on attacking of swine Seneca viruses.

Description

technical field [0001] The invention relates to a Seneca virus isolate, and further relates to an inactivated vaccine for preventing Seneca virus disease prepared by the Seneca virus isolate and an application thereof, belonging to the field of Seneca virus inactivated vaccines. Background technique [0002] In 2002, the Seneca Valley virus strain SVV-001 was accidentally isolated in Gaithersburg, Maryland, USA. Studies have found that it has oncolytic virus properties and is widely used in cancer treatment, and it has been found to have similar characteristics to cardiovirus-related viruses. In 2015, SVV was renamed "Senecavirus A" (SVA), and its genus was named Senecavirus. SVA can cause vesicular disease in pigs characterized by diarrhea, lameness, anorexia, lethargy, vesicles and erosions on the skin, nasal cavity, oral cavity, and coronary artery strips, and its clinical symptoms are similar to those of foot-and-mouth disease virus, vesicular stomatitis virus, Porcine...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00A61K39/125A61P31/14C12R1/93
CPCC12N7/00A61K39/12A61P31/14C12N2770/32021C12N2770/32034A61K2039/5252
Inventor 刘鑫莹杨本孙心李建华李应鹤孙晓峰蒋迪武啸李贽
Owner 哈药集团生物疫苗有限公司
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