Porcine transmissible gastroenteritis virus (PTGEV) and porcine epidemic diarrhea virus (PEDV) dual yolk antibody and preparation method thereof
A technology for porcine epidemic diarrhea and epidemic diarrhea, applied in the direction of antiviral immunoglobulin, antibody, antiviral agent, etc., can solve the problems of unsatisfactory treatment effect, achieve rapid treatment effect, reduce production cost, and simple production procedure Effect
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Embodiment 1
[0038] Example 1 Preparation of Porcine Transmissible Gastroenteritis Virus and Epidemic Diarrhea Virus Double Egg Yolk Antibody
[0039] 1. Preparation of vaccine for immunization:
[0040] 1.1 Proliferation of infectious gastroenteritis virus and preparation of antigen
[0041] Inoculate 3 bottles of PK15 cells with infectious gastroenteritis virus liquid, and inoculate 1ml of virus liquid in each bottle of cells, 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 whole cell monolayer, absorb at 37°C for 40 minutes, shake once every 10 minutes during this period, do not need to suck and discard the virus liquid, directly add the containing 2% fetal bovine serum DMEM (Dulbecco's Modified Eagle Medium) cell maintenance medium 5ml, set up cell control at the same time, at 37 ℃ temperature CO 2 cultured in an incubator. After 24 hours, the maintenance solution was discarded a...
Embodiment 2
[0060] Example 2 Porcine Transmissible Gastroenteritis Virus and Epidemic Diarrhea Virus Duplex Egg Yolk Antibody Titer Detection
[0061] 1. Experimental preparation Porcine transmissible gastroenteritis virus and epidemic diarrhea virus standard positive serum, provided by the Harbin Veterinary Research Institute of the Chinese Academy of Agricultural Sciences; Antigen, prepared by Example 1; Agarose, puncher, 16# needle, scissors , tweezers, alcohol lamp, marker pen, clean plate, water bath.
[0062] 2. Preparation of porcine transmissible gastroenteritis virus (or epidemic diarrhea virus) agar-amplified antigen: use polyethylene glycol (PEG-20000) dry powder to embed a dialysis bag for antigen concentration.
[0063] The treatment method of the dialysis bag: 0.5mol / L EDTA-2Na 1ml, add 10g sodium bicarbonate, add distilled water to 500ml, and adjust the pH value to 8.0; put the dialysis bag in the above liquid and boil for 10min; take out the dialysis bag and clean it with ...
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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