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Preparation method of anti-duck hepatitis virus transfer factor

A technology of duck hepatitis virus and transfer factor, which is applied in the preparation methods of peptides, antiviral agents, microorganism-based methods, etc., can solve the problems of non-specific disease and unstable treatment effect, achieve strong specificity and improve immunity. strength, good effect

Inactive Publication Date: 2016-05-11
TIANJIN HLINTE BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to overcome the disadvantages of preparation in the existing transfer factor technology, lack of specificity for diseases, unstable therapeutic effect, etc., and provide an in vitro immunization method with strong specificity, low cost, high output rate and better effect. Preparation method of anti-duck hepatitis virus specific transfer factor

Method used

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  • Preparation method of anti-duck hepatitis virus transfer factor

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Comparison scheme
Effect test

Embodiment 1

[0023] A kind of in vitro preparation method of anti-duck hepatitis virus specific transfer factor, its steps are as follows:

[0024] (1) Chicken spleen or chicken blood was collected under a sterile environment, and a monolayer of lymphocytes was prepared from the chicken spleen or peripheral blood. Take 30ml of animal whole blood through a sterile vein, add 0.8% heparin solution 0.3m1 for anticoagulation, let it stand for 60 minutes, and use a straw with a rubber ball to absorb all the plasma above the red blood cell layer, especially the white blood cell layer immediately above the red blood cell layer. Dispense into sterilized centrifuge tubes, wash with PBS (pH 7.2) solution repeatedly for 3 times, centrifuge at 1200rpm, then use 40ml of hydrolyzed milk protein containing 30% calf serum for leukocyte culture, mix well and then divide into 100ml cell culture In the bottle, tightly stoppered, placed in a 37°C cell culture incubator, after 3 days, the leukocytes evenly adhe...

Embodiment 2

[0029] A kind of in vitro preparation method of anti-duck hepatitis virus specific transfer factor, its steps are as follows:

[0030] (1) Chicken spleen or chicken blood was collected under a sterile environment, and a monolayer of lymphocytes was prepared from the chicken spleen or peripheral blood. Take 20ml of animal whole blood through a sterile vein, add 0.8% heparin solution 0.3m1 for anticoagulation, let it stand for 30 minutes, and use a straw with a rubber ball to absorb all the plasma above the red blood cell layer, especially the white blood cell layer immediately above the red blood cell layer. Dispense into sterilized centrifuge tubes, wash with PBS (pH 7.2) solution repeatedly for 3 times, centrifuge at 800rpm, then use 40ml of hydrolyzed milk protein containing 30% calf serum for leukocyte culture, mix well and then divide into 100ml cell culture In the bottle, tightly stoppered, placed in a 37°C cell culture incubator, after 2 days, the leukocytes evenly adher...

Embodiment 3

[0035] A kind of in vitro preparation method of anti-duck hepatitis virus specific transfer factor, its steps are as follows:

[0036] (1) Chicken spleen or chicken blood was collected under a sterile environment, and a monolayer of lymphocytes was prepared from the chicken spleen or peripheral blood. Take 25ml of animal whole blood through a sterile vein, add 0.8% heparin solution 0.3m1 for anticoagulation, let it stand for 45 minutes, and use a straw with a rubber ball to absorb all the plasma above the red blood cell layer, especially the white blood cell layer immediately above the red blood cell layer. Dispense into sterilized centrifuge tubes, wash twice with PBS (pH 7.2) solution, centrifuge at 1200rpm, then use 40ml of hydrolyzed milk protein containing 30% calf serum for leukocyte culture, mix well and then divide into 100ml cell culture In the bottle, tightly stoppered, placed in a 37°C cell culture incubator, after 3 days, the leukocytes evenly adhered to the wall, ...

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Abstract

The invention relates to a preparation method of an anti-duck hepatitis virus specific transfer factor. The method includes: inoculating a duck hepatitis virus to the allantoic fluid of a 9-11 days old chicken embryo or duck embryo to prepare a virus antigen, and extracting a duck hepatitis virus antigen; then taking chicken spleen or peripheral blood as the raw material to prepare a lymphocyte monolayer, then using phytohemagglutinin and the duck hepatitis virus to perform induction culture of lymphocyte, thus generating the anti-duck hepatitis virus specific transfer factor in vivo. The specific transfer factor prepared by the method provided by the invention can be used for prevention and treatment of duck hepatitis disease, and also can improve the immunity of ducks. The method provided by the invention has the advantages of strong specificity, low cost, high yield and good effect, etc., and is an ideal preparation method of duck hepatitis virus transfer factor.

Description

technical field [0001] The invention relates to the field of veterinary medicine, in particular to an in vitro preparation method of anti-duck hepatitis virus transfer factor. Background technique [0002] Duck viral hepatitis, referred to as duck hepatitis for short, is a rapidly spreading and highly fatal infectious disease of ducklings caused by duck hepatitis virus. The main features of the disease are hepatic enlargement with spotted hemorrhage, enlarged gallbladder, pale bile, bleeding spots and neurological symptoms. In the initial infection epidemic area, the mortality rate of the disease is very high, which can reach more than 90%. [0003] Duck viral hepatitis mainly occurs in ducklings aged 4 to 20 days. Adult ducks generally do not develop the disease, and chickens and geese cannot naturally develop the disease. The main source of infection of the disease is sick ducks and infected ducks, mainly through digestive tract and respiratory tract infection, poor feed...

Claims

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Application Information

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IPC IPC(8): A61K38/02C07K1/34A61P31/14C12R1/93
Inventor 陈庆忠安同伟高华义
Owner TIANJIN HLINTE BIOTECH CO LTD
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