Preparation method of transfer factor for resisting avian influenza virus

A technology of avian influenza and transfer factor, which is applied in antiviral agents, methods based on microorganisms, biochemical equipment and methods, etc., can solve the problems of non-specificity of diseases and unstable treatment effect, and achieve strong specificity and improved Immunity, good effect

Pending Publication Date: 2022-07-01
DINGZHENG XINXING BIOTECH TIANJIN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] 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 immune method with strong specificity, low cost, high output rate and better effect. Preparation method of anti-avian influenza virus specific transfer factor

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] A preparation method for preparing anti-avian influenza virus specific transfer factor in vitro:

[0018] (1) The chicken spleen was taken in a sterile environment, and the lymphocyte monolayer was prepared from peripheral blood. Take 20ml of animal whole blood from sterile vein, add 0.2ml of 0.8% heparin solution for anticoagulation, let it stand for 40min, and use a pipette with a rubber bulb to absorb all the plasma above the erythrocyte layer, especially the leukocyte layer immediately above the erythrocyte layer. Dispense into sterilized centrifuge tubes, wash 3 times with PBS (pH 7.2), and centrifuge at 1,200 rpm, then use 40 ml of hydrolyzed milk protein containing 30% calf serum for leukocyte culture, mix evenly, and divide into volumes In a 100ml cell culture flask, close the stopper, and place it in a cell culture incubator at 37°C. After 2 days, the leukocytes evenly adhere to the wall, which is a lymphocyte monolayer;

[0019] (2) Subculture the prepared ly...

Embodiment 2

[0023] A preparation method for preparing anti-avian influenza virus specific transfer factor in vitro:

[0024] (1) The chicken spleen was taken in a sterile environment, and the lymphocyte monolayer was prepared from peripheral blood. Take 30ml of animal whole blood from sterile vein, add 0.8% heparin solution 0.2ml for anticoagulation, let it stand for 60min, use a pipette with a rubber bulb 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 3 times with PBS (pH 7.2), centrifuge at 1000 rpm, and then use 40ml of hydrolyzed milk protein containing 30% calf serum for leukocyte culture, mix evenly, and dispense into volume In a 100ml cell culture flask, close the stopper, and place it in a cell culture incubator at 37°C. After 3 days, the white blood cells evenly adhere to the wall, that is, a lymphocyte monolayer is formed;

[0025] (2) Subc...

Embodiment 3

[0029] Detection of the Anti-Avian Influenza Virus Specific Transfer Factor Prepared by the Process of the Invention The following detection is performed on the anti-avian influenza virus transfer factor (hereinafter referred to as "this product") prepared by the process described in Example 1:

[0030] (1) The standard solution is a yellow clear liquid with a pH value between 5.7-7.2;

[0031] (2) Ultraviolet spectrophotometry: this product has a high absorption peak at 280.0 - 290.0nm, and ABS260 / ABS280>2.2;

[0032] (3) Determination of polypeptide content: The polypeptide content of this product was 1.012mg / ml determined by the double-shrinkage method;

[0033] (4) Determination of nucleic acid content: The nucleic acid content of this product was determined by the lichenol method to be 0.472 mg / ml.

[0034] Clinical application

[0035] Test 1

[0036] In October 2019, 5,000 chicken farms infected with avian influenza were selected in Jixian County and treated with the...

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Abstract

The invention relates to a method for preparing a specific transfer factor for resisting avian influenza virus in vitro. Chicken spleen or peripheral blood is used as a raw material to prepare a lymphocyte monolayer, then phytohemagglutinin and avian influenza virus are used for inducing and culturing lymphocytes, and the avian influenza virus resistant specific transfer factor is produced in vitro. The prepared specific transfer factor can be used for preventing and treating avian influenza, and can improve the immunity of chickens. The preparation method has the advantages of high specificity, low cost, good effect and the like, and is an ideal preparation method of the transfer factor for resisting the avian influenza virus.

Description

technical field [0001] The invention relates to the field of veterinary medicine, and more specifically, relates to a preparation method of an anti-avian influenza virus transfer factor. Background technique [0002] It is an acute infectious disease caused by a subtype of influenza A virus (also known as avian influenza virus), which can also infect humans. . Symptoms of human infection are mainly high fever, cough, runny nose, myalgia, etc. Most of them are accompanied by severe pneumonia. In severe cases, heart, kidney and other organ failure leads to death, and the mortality rate is very high. Usually people are infected with avian influenza. The mortality rate is about 33%. The disease can be transmitted through a variety of routes such as digestive tract, respiratory tract, skin injury and conjunctiva, and the movement of people and vehicles between regions is an important way to spread the disease. In recent years, a new subtype of influenza virus H has also appear...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/17C12N5/078C12N7/02A61P31/16A61P11/00A61P37/04C12R1/93
CPCA61K35/17A61P31/16A61P11/00A61P37/04C12N5/0634C12N7/00C12N2760/16151
Inventor 刘鼎阔刘芳李源
Owner DINGZHENG XINXING BIOTECH TIANJIN
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