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Porcine epidemic diarrhea virus variant strain and inactivated vaccine and application thereof

A porcine epidemic diarrhea and virus mutation technology, applied to vaccines, viruses, antiviral agents, etc., to achieve good immune effect and good safety

Active Publication Date: 2018-09-04
黑龙江正康生物技术股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, there is an urgent need to develop an inactivated vaccine with good safety and immune effect on porcine epidemic diarrhea variant strains, so as not to cause serious economic losses to farmers

Method used

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  • Porcine epidemic diarrhea virus variant strain and inactivated vaccine and application thereof
  • Porcine epidemic diarrhea virus variant strain and inactivated vaccine and application thereof
  • Porcine epidemic diarrhea virus variant strain and inactivated vaccine and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 Isolation and Identification of Porcine Epidemic Diarrhea Virus Strains

[0030] 1. Cells and disease materials

[0031] Vero cells were purchased from the China Veterinary Drug Administration; disease materials were collected from the small intestine, contents of the small intestine, and feces of suckling piglets on the verge of death in a large-scale pig farm in Hebei Province.

[0032] Grind the disease material with a grinder, freeze and thaw repeatedly 3 times, mix it with PBS (0.1mol / L, pH 7.2) at 1:5 (V / V), centrifuge at 3000r / min for 15min, take the supernatant, and pass through 0.22 After sterilizing with a μm filter membrane, store at -70°C for later use.

[0033] 2. Virus isolation

[0034] Put the above-mentioned disease material into the Vero cell culture that forms a monolayer according to the inoculum amount of 10% of the virus culture solution, place it at 37°C for 1 hour, replace it with DMEM culture solution containing 2% newborn bovine ser...

Embodiment 2

[0053] Example 2 Preparation of porcine epidemic diarrhea disease inactivated vaccine and evaluation of its safety and immune efficacy 1. Preparation of virus liquid for vaccine production

[0054] Inject the porcine epidemic diarrhea virus PEDV-HB strain isolated in Example 1 into a monolayer of Vero cells by 1% inoculation amount, place 37°C for rotary culture, and when the cytopathic changes reach 80%, harvest the toxic cells for culture After 3 times of freezing and thawing, the virus liquid was harvested.

[0055] 2. Preparation of virus inactivated vaccine

[0056] Add β-propiolactone with a final concentration of 0.2% (w / v, g / ml) to the above-mentioned harvested virus liquid, and inactivate at 30° C. for 36 hours. Mix the porcine epidemic diarrhea virus PEDV-HB strain virus liquid that has passed the inactivation test according to the ratio (v / v) of Montanide ISA 15A VG adjuvant = 9:1, first add the water phase to the emulsification tank and stir slowly , and then slo...

Embodiment 3

[0066] The comparative test of the immune effectiveness of embodiment 3 porcine epidemic diarrhea mutant strain inactivated vaccine and porcine epidemic diarrhea classical strain inactivated vaccine

[0067] 1. Inactivated vaccine against variant strain of porcine epidemic diarrhea

[0068] The porcine epidemic diarrhea variant inactivated vaccine prepared in Example 2.

[0069] 2. Duplex inactivated vaccine against porcine transmissible gastroenteritis and porcine epidemic diarrhea

[0070] Since there is no single vaccine against porcine epidemic diarrhea in the market, a commercially available dual vaccine was selected for the test. The vaccine is a product of Harbin Veken Biotechnology Development Co., Ltd., batch number 20150804. The antigen used for the production of the double vaccine is the classic strain of porcine epidemic diarrhea CV777.

[0071] 3. Porcine epidemic diarrhea classic strain CV777

[0072] For the determination of serum neutralizing antibodies.

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Abstract

The invention discloses a porcine epidemic diarrhea virus variant strain and an inactivated vaccine and application thereof. The invention discloses an isolated porcine epidemic diarrhea virus variantstrain PEDV-HB with the preservation number of CGMCC No. 14889. The invention further discloses a use of the variant strain PEDV-HB in preparation of a vaccine for preventing porcine epidemic diarrhea. The invention also discloses a vaccine composition for preventing porcine epidemic diarrhea. The vaccine composition comprises a prophylactically effective amount of the inactivated porcine epidemic diarrhea virus variant strain PEDV-HB and pharmaceutically acceptable adjuvants. The inactivated vaccine prepared from the porcine epidemic diarrhea virus variant strain PEDV-HB has good safety andgood immune effects, and can better prevent and control porcine epidemic diarrhea. The isolated porcine epidemic diarrhea virus strain PEDV-HB can be used as a candidate vaccine strain for preventionand control of porcine epidemic diarrhea.

Description

technical field [0001] The present invention relates to an isolated variant strain of porcine epidemic diarrhea virus, and also relates to an inactivated vaccine prepared from the variant strain of porcine epidemic diarrhea virus and its application in preventing porcine epidemic diarrhea disease, which belongs to porcine epidemic diarrhea virus Isolation and application fields of mutant strains. Background technique [0002] Porcine epidemic diarrhea is a contagious intestinal infectious disease caused by porcine epidemic diarrhea virus (PEDV, Porcine Epidemic Diarrheavirus). The disease only occurs in pigs, and pigs of all ages can be infected, but it causes the most serious harm to suckling piglets, and the incidence rate of sows is about 15% to 90%. Clinical features include vomiting, diarrhea, and dehydration. The severity of symptoms varies with age. The younger the symptoms, the heavier the symptoms. Newborn piglets (within 1 week of age) will die of severe dehydrat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00A61K39/215A61P31/14
CPCA61K39/12C12N7/00A61K2039/5252A61K2039/552C12N2770/20034
Inventor 杨万秋王东升张明波
Owner 黑龙江正康生物技术股份有限公司
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