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Preparation method of oral vaccine used for treating porcine epidemic diarrhea virus

A porcine epidemic diarrhea and oral vaccine technology, which is applied in the field of preparation of new oral vaccines for effective prevention and control of porcine epidemic diarrhea virus, to achieve the effects of avoiding blindness, stimulating mucosal immune response, and reducing stress

Active Publication Date: 2018-11-30
WENS FOODSTUFF GRP CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In order to overcome the shortcomings of the existing vaccines that cannot avoid gastric acid damage and cannot well stimulate pig intestinal tract to produce sufficient PEDV-specific mucosal immunity, the present invention prepares a new type of PEDV that can tolerate gastric acid and effectively stimulate pig intestinal mucosal immunity. Oral vaccine lays the foundation for the prevention and control of PED in production, and also provides new ideas for the prevention and control of other porcine intestinal infectious diseases

Method used

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  • Preparation method of oral vaccine used for treating porcine epidemic diarrhea virus
  • Preparation method of oral vaccine used for treating porcine epidemic diarrhea virus
  • Preparation method of oral vaccine used for treating porcine epidemic diarrhea virus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Embodiment 1 The preparation of a novel oral vaccine for the effective prevention and control of porcine epidemic diarrhea virus by the present invention

[0043] Including the following actions:

[0044] 1. Virus freeze-drying: the attenuated strain of porcine epidemic diarrhea virus (PEDV) that African green monkey kidney (Vero) cells have cultivated in large quantities is filtered with 4 layers of sterile gauze and with an equal volume of aseptic freeze-drying protectant (containing 10% skimmed milk powder, 5% sucrose) and stir well. Next, turn on the lyophilizer for pre-freezing. When the temperature of the plate drops to -50°C, put the mixed virus solution into the tray of the lyophilizer, and keep it at -50°C for 2 hours after entering the box; Vacuum, open the butterfly valve, and turn on the Roots pump; reduce the vacuum degree of the chamber to 20pa and maintain it for 1h; after 1h, heat for 200min to raise the sample temperature to -7°C and maintain it for mo...

Embodiment 2

[0048] Example 2 Vaccine function test prepared by the method of the present invention

[0049] 2.1 In vitro acid test:

[0050] Weigh 2g NaCl, 3.2g pepsin (12000U) and fully dissolve in 900ml distilled water to prepare artificial gastric juice, then add concentrated HCL to adjust the pH value to 1.2, and dilute to 1L. The prepared PEDV new oral vaccine was placed in a small cup of an intelligent dissolution tester by the small cup method (paddle method), then 100 mL of artificial gastric juice was added, and stirred at 37° C. and 100 r / m for 2 hours. After 2 hours, the artificial gastric juice was poured out, washed with pure water and transferred to pH 6.8 DMEM cell culture medium for dissolution and live virus titer determination. The results are shown in Figure 4 , in which PEDV freeze-dried powder has not been treated with artificial gastric acid TCID 50 =10 6.5 , treated with artificial gastric acid TCID 50 = 0; TCID of PEDV oral vaccine without artificial gastric a...

Embodiment 3

[0072] Refer to the preparation method of the novel oral vaccine for the prevention and control of porcine epidemic diarrhea virus in Example 1, the difference is that in step 2, the virus freeze-dried powder is fully mixed with cornstarch, lactose, dextrin, and mannitol of equal quality, and the other steps Operation is the same as embodiment one. Observe the state of each freeze-dried powder mixture and the subsequent process conditions.

[0073] In the solution of the present invention, the freeze-dried powder contains sucrose milk powder, which is easy to absorb moisture and become lumpy due to moisture. The cornstarch is very fine and has strong hygroscopicity. Mixing with the freeze-dried powder can play the role of dilution and moisture absorption, and keep the flow of the freeze-dried powder. State, and in the enteric coating stage, starch can act as a disintegrant to accelerate the release of the virus in the intestine. And the yield of powdering with cornstarch is h...

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Abstract

The invention belongs to the field of veterinary biological technology application, and more specifically relates to a preparation method of an oral vaccine used for treating porcine epidemic diarrheavirus (PEDV). The preparation method comprises following steps: a virus low virulent strain is subjected to freeze drying; obtained virus freeze-dried powder is subjected to centrifugation granulation so as to obtain pellets loaded with the virus low virulent strain; the pellets are coated with isolating coating; the pellets coated with isolating coating is coated with enteric coating so as to obtain the oral vaccine. The PEDV novel oral vaccine is capable of resisting hydrochloric acid in gastric juice, and stimulating porcine intestinal tract mucous membrane immunization, is used for prevention and control of PED in production, and can be mixed with forage for feeding of pigs; operation is convenient; stress and feedback virus dispersion risks caused by other immunization methods are reduced greatly; and the safety is better.

Description

technical field [0001] The invention belongs to the application field of veterinary biotechnology, and in particular relates to a preparation method of a novel oral vaccine for effectively preventing and controlling porcine epidemic diarrhea virus. Background technique [0002] Porcine epidemic diarrhea (Porcine epidemic diarrhea, PED) is an acute and highly contagious disease of pigs caused by porcine epidemic diarrhea virus (Porcine epidemic diarrhea virus, PEDV). Newborn piglets infected with PEDV developed diarrhea, dehydration, and vomiting, and had a high mortality rate. In recent years, PED has been widely prevalent in the world, and outbreaks have occurred in China, the United States, South Korea, Canada and other countries, seriously endangering the development of the global pig industry. PED occurs and is prevalent in 29 provinces and autonomous regions in my country, which can lead to a mortality rate of up to 100% in newborn piglets, and has caused huge economic...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/215A61K9/52A61K47/32A61K47/26A61P31/14
CPCA61K39/12A61P31/14A61K9/5026A61K9/5078A61K2039/5254A61K2039/552A61K2039/575C12N2770/20034
Inventor 温志芬李薇周庆丰徐志超吴云燕曹永长
Owner WENS FOODSTUFF GRP CO LTD
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