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Method achieving homeomorphic cultivation of Newcastle disease and infectious bronchitis virus

A technology for chicken Newcastle disease virus and bronchitis, which is applied to viruses/phages, biochemical equipment and methods, microorganisms, etc., can solve the problems of large batch differences, poor stability of homoembryonic culture, and lack of broad-spectrum. Batch-to-batch variation, improved virus resistance, improved stability effects

Pending Publication Date: 2016-01-06
浙江美保龙生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the effect of polysaccharides in traditional Chinese medicine does not have a broad spectrum, and the synergistic effect of one or several polysaccharides has specific active effects.
[0009] Based on the above technical background, the present invention proposes a method for homeomorphic culture, specifically a method especially suitable for homeomorphic culture of chicken Newcastle disease and infectious bronchitis virus, which solves the problem of homeomorphic culture of Newcastle disease virus and chicken infectious bronchitis virus. Improve the stability, yield and virus titer of the two viruses due to poor culture stability and large batch-to-batch differences

Method used

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  • Method achieving homeomorphic cultivation of Newcastle disease and infectious bronchitis virus
  • Method achieving homeomorphic cultivation of Newcastle disease and infectious bronchitis virus
  • Method achieving homeomorphic cultivation of Newcastle disease and infectious bronchitis virus

Examples

Experimental program
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Effect test

Embodiment 1

[0039] A chicken Newcastle disease virus dilution, the virus dilution contains per 1000ml of phosphate buffer: amino acid combination 3.5g (wherein phenylalanine 0.7g, cystine 1.4g, leucine 1.4g), vitamin combination 6g (wherein Vitamin B21.2g, vitamin D3g, vitamin E1.8g), compound traditional Chinese medicine polysaccharide 50g, sodium selenite 3mg, the pH of the diluent is 6.8.

[0040] A kind of preparation method of chicken Newcastle disease virus dilution, its steps are:

[0041] (1) Preparation of phosphate buffer solution: Accurately weigh 8 g of sodium chloride, 0.2 g of potassium chloride, 1.15 g of disodium hydrogen phosphate, 0.2 g of potassium dihydrogen phosphate, and 0.1 g of calcium chloride according to the above formula of phosphate buffer solution. Dissolve 0.1 g of magnesium chloride containing 6 crystal waters in 1000 ml of double distilled water, pass through a 0.22 μm filter membrane under aseptic conditions, and filter and sterilize for later use.

[00...

Embodiment 2

[0062] A chicken Newcastle disease virus dilution, this virus dilution is every 1000ml phosphate buffer (this phosphate buffer is made of sodium chloride 6g, potassium chloride 0.05g, disodium hydrogen phosphate 1g, potassium dihydrogen phosphate 0.05g, calcium chloride 0.05 g, 0.05g magnesium chloride containing 6 crystal waters dissolved in 1000ml double distilled water) contains: amino acid combination 4g (including phenylalanine 1g, cystine 1.6g, leucine 1.4g), vitamin combination 7g ( Among them, vitamin B21.2g, vitamin D3g, vitamin E2.8g), compound Chinese medicine polysaccharide 40g (extracted from 20 parts of red peony root, 5 parts of salvia miltiorrhiza, 10 parts of Bai Wei, 5 parts of pokeweed and 10 parts of Cordyceps), selenium 5 mg of sodium bicarbonate, the pH of the diluent is 6.9.

[0063] A kind of preparation method of chicken Newcastle disease virus dilution, its step is the same as embodiment 1.

[0064] A chicken infectious bronchitis virus dilution, the...

Embodiment 3

[0067] A chicken Newcastle disease virus dilution, this virus dilution is every 1000ml phosphate buffer (this phosphate buffer is made of sodium chloride 10g, potassium chloride 0.5g, disodium hydrogen phosphate 1.2g, potassium dihydrogen phosphate 0.5g, calcium chloride 0.2g, 0.2g of magnesium chloride containing 6 crystal water dissolved in 1000ml of double distilled water) contains: amino acid combination 3g (including phenylalanine 1.5g, cystine 0.6g, leucine 0.9g), vitamin combination 10g (including 24g of vitamin B, 2g of vitamin D, and 4g of vitamin E), 100g of compound Chinese herbal polysaccharides (extracted from 40 parts of red peony root, 20 parts of salvia miltiorrhiza, 30 parts of Baiwei, 20 parts of pokeweed and 30 parts of Cordyceps), sodium selenite 10mg, the pH of the diluent is 6.8.

[0068] A kind of preparation method of chicken Newcastle disease virus dilution, its step is the same as embodiment 1.

[0069] A chicken infectious bronchitis virus dilution,...

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Abstract

The invention provides a method achieving homeomorphic cultivation of a Newcastle disease and an infectious bronchitis virus, and belongs to the technical field of biological products for veterinary. The method comprises the steps that Newcastle disease virus diluent is adopted to dilute a Newcastle disease virus for 1000-5000 times, and infectious bronchitis virus diluent is adopted to dilute the infectious bronchitis virus for 5000-10000 times; the Newcastle disease virus and the infectious bronchitis virus are incubated at 4 DEG C for 30-60 min and then evenly mixed according to the proportion of 1 to 1, chick embryos with 9-10 day age are inoculated, each embryo is inoculated for 0.1-0.15 ml, and cultivation is continuously conducted for 96 h at the temperature ranging from 36 DEG C to 37 DEG C; chicken embryos which die before 48 hours are discarded, dead and live embryo liquid with 48-96 h is obtained, and mixed virus liquid of the Newcastle disease virus and an infectious bronchitis virus is obtained. According to the virus liquid which is cultivated and obtained through the method, the stability of homeomorphic cultivation is improved, the differences between batches are greatly reduced, and high-quality antigen liquid is provided for the subsequent vaccine production process.

Description

technical field [0001] The invention belongs to the technical field of veterinary biological products, and in particular relates to a homeomorphic culture method for chicken Newcastle disease and infectious bronchitis virus. Background technique [0002] Newcastle disease (ND) is a severe viral infectious disease caused by Newcastle disease virus (NDV). It is one of the most serious poultry infectious diseases in the world today and the most harmful of all chicken diseases in my country It is classified as a class A infectious disease by the International Office of Epizootics (OIE). [0003] Chicken infectious bronchitis (IB) is an acute, highly contagious viral disease of chickens caused by infectious bronchitis virus (IBV), characterized by tracheal rales, coughing, and sneezing. One of the main diseases that harm the poultry industry. There are many serotypes of chicken infectious bronchitis, and more than 30 serotypes have been reported so far. Different serotype strain...

Claims

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Application Information

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IPC IPC(8): C12N7/00
Inventor 沈建军张秀文李阳
Owner 浙江美保龙生物技术有限公司