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Porcine transmissible gastroenteritis virus (PTGEV) and porcine epidemic diarrhea virus (PEDV) dual yolk antibody and preparation method thereof

一种猪流行性腹泻、流行性腹泻的技术，应用在抗病毒免疫球蛋白、抗体、抗病毒剂等方向，能够解决治疗效果不理想等问题，达到治疗效果迅速、疗效显著、生产程序简单的效果

Active Publication Date: 2013-07-03
PULIKE BIOLOGICAL ENG INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since there is no specific drug for viral diseases, clinical treatment mainly adopts symptomatic treatment and antibiotics to prevent complications, and the treatment effect is not ideal

Method used

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  • Porcine transmissible gastroenteritis virus (PTGEV) and porcine epidemic diarrhea virus (PEDV) dual yolk antibody and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1 Preparation of swine transmissible gastroenteritis virus and epidemic diarrhea virus double yolk antibody

[0039] 1. Preparation of vaccines for immunization:

[0040] 1.1 Transmissible gastroenteritis virus proliferation and antigen preparation

[0041] Inoculate 3 bottles of PK15 cells with infectious gastroenteritis virus solution, each bottle of cells is inoculated with 1ml of virus solution, and the bottom area of ​​the bottle is 25cm 2 , cover the bottle cap, turn the horizontal bottle tube gently to make the virus liquid fully contact with the entire cell monolayer, adsorb at 37°C for 40min, shake once every 10min during this period, no need to aspirate and discard the virus solution, directly add containing 5ml of DMEM (Dulbecco's Modified Eagle Medium) cell maintenance medium containing 2% fetal bovine serum, and a cell control was established at the same time, at 37°C with CO 2 Cultivated in an incubator. After 24 hours, the maintenance solution ...

Embodiment 2

[0060] Example 2 Detection of double yolk antibody titers of porcine transmissible gastroenteritis virus and epidemic diarrhea virus

[0061] 1. Experimental preparation Standard positive serum of porcine transmissible gastroenteritis virus and epidemic diarrhea virus, provided by Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences; antigen, prepared from Example 1; agarose, punch, 16# needle, scissors , tweezers, alcohol lamp, marker, clean plate, water bath.

[0062] 2. Preparation of swine transmissible gastroenteritis virus (or epidemic diarrhea virus) agar-expanded antigen: the dialysis bag was embedded with polyethylene glycol (PEG-20000) dry powder for antigen concentration.

[0063] The treatment method of the dialysis bag: add 10g of sodium bicarbonate to 1ml of 0.5mol / L EDTA-2Na, add distilled water to 500ml, and adjust the pH value to 8.0; boil the dialysis bag in the above liquid for 10min; take out the dialysis bag and clean it with dist...

Embodiment 3

[0079] Example 3 Therapeutic Test of Porcine Transmissible Gastroenteritis Virus and Epidemic Diarrhea Virus Dual Egg Yolk Antibody

[0080] 16 1-day-old piglets that did not eat colostrum were taken, and the temperature of the experimental animal room was controlled at 25-30°C. Artificial lactation was adopted, and milk was fed once every 2 hours. After feeding and observation for 1 day, they were randomly divided into 4 groups. They are: treatment group (4 animals), drug attack group (4 animals), drug treatment group (4 animals), and no treatment group (4 animals). The viruses used for the challenge are ultrafiltered porcine transmissible gastroenteritis virus and epidemic diarrhea virus, which are diluted 10 times with a phosphate buffer solution with a concentration of 0.01mol / L and a pH of 7.2, and are directly orally administered. 5 times of that, the dose of each piglet was the same, and the diarrhea situation of the piglets after the challenge was observed. When the p...

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Abstract

The invention discloses a method for preparing a double yolk antibody of a porcine transmissible gastroenteritis virus and porcine epidemic diarrhea virus. The method comprises the following steps of: performing porcine transmissible gastroenteritis virus multiplication on porcine kidney cells (PK15); performing porcine epidemic diarrhea virus multiplication on African green monkey kidney cells (Vero); emulsifying the two cell cultures used as antigen with an oil emulsion adjuvant to prepare immunogen, namely, mixing the two kinds of viruses in a ratio of (1-3):(1-3) to prepare the immunogen; immunizing non-immunologic laying hens; and obtaining the double yolk antibody which can prevent and treat porcine transmissible gastroenteritis and porcine epidemic diarrhea based on the collection and purification of the yolk. When the double yolk antibody is used for curing experimental pigs, the clinical symptoms in the experiment are obviously reduced compared with a control group, and the death rate of the experimental group is obviously lower than that of the control group. The double yolk antibody has obvious preventing and treating functions when applied in a pig farm with high incidence rate of the porcine transmissible gastroenteritis and the porcine epidemic diarrhea.

Description

technical field [0001] The invention belongs to the technical field of veterinary biological products, in particular to a porcine infectious gastroenteritis virus, a porcine epidemic diarrhea virus double yolk antibody and a preparation method thereof. Background technique [0002] Porcine infectious gastroenteritis and porcine epidemic diarrhea are two important viral infectious diseases that have been confirmed to cause diarrhea in pigs. These two diseases mainly have a greater impact on piglets and are the main viral diarrheal diseases that cause piglets to die. These two diseases are often co-infected, and are very similar in epidemiology, clinical symptoms, and pathological anatomical changes. It is difficult to distinguish which virus infection is, and the onset is rapid, which can spread within a few hours after the onset. The whole herd, mainly causing a large number of deaths of lactating piglets, caused huge losses to pig production. [0003] Porcine Transmissibl...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/10C07K16/02A61K39/42A61P1/00A61P1/04A61P1/12A61P31/14
Inventor 张许科孙进忠乔荣岑
Owner PULIKE BIOLOGICAL ENG INC
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