Method for preparing double yolk antibody of porcine transmissible gastroenteritis virus and porcine epidemic diarrhea virus

A technology for porcine epidemic diarrhea and epidemic diarrhea, which is applied in the direction of antiviral immunoglobulin and egg immunoglobulin, etc., can solve problems such as unsatisfactory treatment effect, and achieve highly specific and targeted effects.

Inactive Publication Date: 2010-09-29
PU LIKE BIO ENG
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Problems solved by technology

Since there is no specific drug for viral diseases, clinical treatment mainly adopts sympt...

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  • Method for preparing double yolk antibody of porcine transmissible gastroenteritis virus and porcine epidemic diarrhea virus

Examples

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Effect test

Embodiment 1

[0034] Propagation and Antigen Preparation of Porcine Transmissible Gastroenteritis Virus (TGEV) and Porcine Epidemic Diarrhea Virus (PEDV)

[0035]1. TGEV virus proliferation and antigen preparation

[0036] Inoculate 3 bottles of PK15 cells with infectious gastroenteritis virus liquid (bottle bottom area 25cm 2 ), each bottle of cells was inoculated with 1ml of virus solution, capped the bottle cap, and the horizontal bottle tube was rotated gently to make the virus solution fully contact with the entire cell monolayer, and adsorbed at 37°C for 40 minutes, during which it was shaken every 10 minutes, no need to absorb and discard For the virus solution, directly add 5ml of DMEM cell maintenance medium containing 2% fetal bovine serum, and set up a cell control at the same time, at 37°C CO 2 cultured in an incubator. After 24 hours, the maintenance solution was discarded and the medium was changed. Observe the cell lesions every day. If the infected cell lesions (CPE) are ...

Embodiment 2

[0039] Example 2 Preparation of TGEV-PEDV Double Egg Yolk Antibody

[0040] 1. Immunogen Preparation

[0041] After the harvested TGEV antigen and PEDV antigen were mixed in a ratio of 1:1, 0.1% of the total amount was added to formaldehyde solution (40% aqueous solution), and inactivated at 37° C. for 24 hours as the immunogen for the first immunization. Mix 96 parts of the above-mentioned inactivated antigen plus 4 parts of Tween-80 as the water phase (preparation before use); preparation of the oil phase: 94 parts of white oil + 6 parts of Span-80, and then press 0.015g / ml Add aluminum stearate in an appropriate amount, heat and mix in a water bath, autoclave at 121°C for 30 minutes, and set aside; emulsification: first add the oil phase, pre-stir at 3000 rpm for 5 minutes, then slowly add the water phase, shake while adding, and stir for 30 seconds. Adjust the rotation speed to 12000rpm, stir for 5 minutes, and put it at 4°C for later use. Take 10ml of emulsified inactiv...

Embodiment 3

[0078] Example 3 TGEV-PEDV dual egg yolk antibody treatment test

[0079]Take 16 piglets who did not eat colostrum and have no history of TGEV and PEDV at the age of 1 day. 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. They were divided into 4 groups, namely: treatment group (4 animals), treatment group (4 animals), no treatment group (4 animals), no treatment group (4 animals), and no treatment group (4 animals). The viruses used for the challenge were ultrafiltered TGEV and PEDV diluted 10 times with 0.01mol / L PBS (pH7.4) and directly orally administered. Diarrhea in piglets after challenge. When the piglets first started to have diarrhea symptoms, the treatment group was fed TGEV-PEDV double egg yolk antibody, the egg yolk antibody used in the treatment was a purified sample, and the titer of the egg yolk antibody determined by the agar diffusion test was 1:64. The non-trea...

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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, and in particular relates to a double egg yolk antibody against porcine transmissible gastroenteritis virus and porcine epidemic diarrhea virus and a preparation method thereof. Background technique [0002] Porcine transmissible 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 diarrhea diseases that cause piglet death. These two diseases are often co-infected, and are very similar in epidemiology, clinical symptoms, and pathological and anatomical changes. The whole group mainly causes a large number of deaths of lactating piglets, causing huge losses to pig production. [0003] Porcine Transmissible Gastroenteritis (TGE) is caused by porcine transmissible gastroenteritis virus (Porcine Transmissib...

Claims

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Application Information

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IPC IPC(8): C07K16/10C07K16/02
Inventor 张许科孙进忠乔荣岑陶家权郭丽霞张海洋习向锋李三
Owner PU LIKE BIO ENG
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