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Duck leukocyte interferon for treating duck viral disease

A leukocyte interferon, viral disease technology, applied in the direction of cytokine/lymphokine/interferon, interferon, antiviral agent, etc., can solve the problem of different sensitivity of interferon, achieve huge market prospects, low dosage , the effect of high preparation cost

Inactive Publication Date: 2010-12-08
TIANJIN RINGPU BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] 2. Interferon is species-specific, and human interferon is generally only used to treat human diseases
[0007] 3. Different cell types, different animals, and different virus infection types have different sensitivities to interferon
There is no report about the preparation of duck leukocyte interferon by using duck blood

Method used

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  • Duck leukocyte interferon for treating duck viral disease
  • Duck leukocyte interferon for treating duck viral disease

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Embodiment 1: a kind of preparation method of duck leukocyte interferon, concrete steps are as follows:

[0021] (1) Preparation of duck serum: aseptically collect duck blood, place it in a refrigerator at 4°C for natural precipitation, and use 0.2% CaCl 2 Process the plasma, remove fibrin, filter and sterilize at 0.22 μm, store at -20°C for later use, the dosage is 4-8%;

[0022] (2) Preparation of white blood cells: Aseptically collect healthy duck blood, add 100 μg / ml double antibody and appropriate amount of citric acid anticoagulant, place in a refrigerator at 4°C; centrifuge at 4000 rpm for 30 minutes, collect middle white blood cells, sallow layer, and wash with PBS Centrifuge again, collect the precipitated white blood cells, and dilute the white blood cell precipitate with DMEM culture solution added with 100 μg / ml double antibody and the above-mentioned duck serum, so that the white blood cell concentration is diluted to 1×10 6 / ml;

[0023] (3) Add human le...

Embodiment 2

[0026] Embodiment 2: a kind of method for duck leukocyte interferon antiviral activity detection, concrete steps are as follows:

[0027] (1) Remove the limbs, head and viscera from the 10-day-old SPF duck embryo, rinse twice with PBS with 100 μg / ml double antibody, wash away the blood, cut into pieces, add 2 times the volume of trypsin to digest, 37 ° C for 10 min about;

[0028] (2) Pour off the trypsin when the tissue is cut into flocs, add DMEM culture solution, and pipette repeatedly;

[0029] (3) Filtrate with gauze after beating evenly, remove undigested bulk and impurities, and prepare DEF;

[0030] (4) Take a drop of filtrate to check the cell density on a cell counting plate, and use 1×10 4 Cells / well density Spread DEF on a 96-well plate, 100 μl per well;

[0031] (5) After the cells grow into a monolayer, add 100 μl of 10-fold serial dilution of duck leukocyte interferon, 37 ° C, CO 2 , the effect is about 24h;

[0032] (6) Receive VSV virus 1000 TCID 50 , 37...

Embodiment 3

[0039] Embodiment 3: the application of this duck leukocyte interferon pair on ducks

[0040] For the duck leukocyte interferon prepared by the process described in Example 1, the following research is carried out according to the requirements of the clinical trial research data of veterinary biological products issued by the Ministry of Agriculture Announcement No. 442:

[0041] 1. Experimental animals: 80 20-day-old healthy ducks were fed in a conventional way.

[0042] 2. Toxic challenge: 60 duck plague attenuated to the same amount as the challenge, all caused the disease.

[0043]3. Test method: When the symptoms appear, conduct a clinical treatment test immediately. Randomly divided into 4 groups, 20 in each group. The dose of low-dose duck leukocyte interferon solution is 20IU / kg; the dose of medium-dose duck leukocyte interferon solution is 60IU / kg; the dose of high-dose duck leukocyte interferon solution is 100IU / kg, the blank group does not administer, injects, all...

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Abstract

The invention discloses a preparation and activity detection method for duck leukocyte interferon for treating duck viral disease. Duck source newcastle disease virus is used as an inducer for inducing healthy duck leukocyte. The method comprises the processes of raw material preparation, mixing, water bath stirring culture, centrifugation and harvesting, virus inactivation, sterilization, anti-virus activity detection and the like. The fresh complete blood of a duck is successfully used as a raw material, the preparation process is simple and convenient, and the product quality is reliable. After the duck leukocyte interferon is used for a duck body, the duck leukocyte interferon has the characteristics of good treatment effect, safety, reliability, no toxic or side effect and the like. The duck leukocyte interferon for broad spectrum virus resistance has the function of treating the viral diseases such as duck plague, duck viral hepatitis, duck source newcastle disease, duck source egg drop syndrome and the like. The duck leukocyte interferon is used for the animal body, and has the advantages of good treatment effect, safety, reliability, low using amount, no toxic or side effect, low cost and the like.

Description

technical field [0001] The invention belongs to the technical field of veterinary medicine, and relates to a preparation method of duck leukocyte interferon for treating duck viral diseases and a measurement of its antiviral activity. Background technique [0002] With the rapid development of the national economy and the scientific adjustment of the agricultural structure, the duck industry has become a fast-growing industry of animal husbandry. Under the background of the rapid development of duck industry, the mobility of ducks has greatly increased, the degree of intensive production of ducks has been further improved, and the impact of duck diseases on duck production has become more and more prominent. How to effectively prevent duck diseases and reduce economic losses has become one of the issues that everyone is very concerned about. At present, the occurrence of duck viral diseases is mainly controlled by means of epidemic prevention. In recent years, various stud...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/555A61P31/12
Inventor 王小娥
Owner TIANJIN RINGPU BIO TECH
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