Method for preparing infectious bronchitis viral antigen by adopting non-immune eggs

A technology for chicken infectiousness and bronchitis, applied in the direction of virus antigen components, biochemical equipment and methods, viruses, etc., can solve the problems of high vaccine production costs, reduce vaccine research and production costs, improve quality and output, and have application prospects broad effect

Active Publication Date: 2019-11-05
商丘美兰生物工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The object of the present invention is to be difficult to determine factors such as temperature, humidity and chicken embryo harvesting time in the hatching process of chicken embryo, and adopt SPF chicken embryo

Method used

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  • Method for preparing infectious bronchitis viral antigen by adopting non-immune eggs
  • Method for preparing infectious bronchitis viral antigen by adopting non-immune eggs
  • Method for preparing infectious bronchitis viral antigen by adopting non-immune eggs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Embodiment one process condition is determined

[0021] 1.1. Determine the temperature and humidity in the pre-hatching conditions

[0022] Appropriate temperature and humidity conditions can promote the development of chicken embryos. In view of the 21-day development process of chicken embryos, the fetus sinks into the yolk on the 7th day, and the volume of the yolk reaches the largest. . Using the orthogonal test method, the temperature screening is divided into 4 conditions of 37.8°C, 38.0°C, 38.2°C, and 38.4°C. The humidity screening is divided into two schemes. The first scheme is to select 65%, 70 %, 75% in total 3 humidity conditions; the second option is based on the developmental characteristics of chicken embryos, choose 55%, 60%, 65% in total 3 humidity conditions on the 1st to 7th day, and use 65% on the 8th to 10th day , 70%, 75% total 3 humidity conditions; chicken infectious bronchitis non-immune eggs 480, 10 eggs in each group, calculate the average h...

Embodiment 2

[0050] 2018.09.10 Purchased 40,000 non-immune chicken eggs from Liaocheng, Shandong. The HI (hemagglutination inhibition test) of the eggs was sampled. Control the temperature at 38.2°C, hatch for 10 days, the relative humidity is 60% on the 1st to 7th day, and the relative humidity is 70% on the 8th to 10th day; 5.0 EID 50 The inoculation dose per piece is used to inoculate the chicken infectious bronchitis virus H120 strain in the allantoic cavity inoculation method; the post-hatch hatching conditions are controlled at a temperature of 37.8 ° C and a relative humidity of 70%; 24 hours after inoculation, the dead embryos are removed and the virus is inoculated. After 36 hours, the chicken embryos were placed in a 2-8°C refrigerator, and the virus allantoic fluid was harvested after about 12 hours of refrigeration. The virus content of this batch of chicken infectious bronchitis virus antigens measured is 10 7.3 EID 50 / 0.1mL, the average amount of allantoic fluid harvested...

Embodiment 3

[0052] 2018.10.22 Purchased 52,000 non-immune chicken eggs from Shangqiu, Henan Province. The HI (hemagglutination inhibition test) of the eggs was sampled, and the antibody titer was lower than 1:4; 2018.10.23 Incubation, pre-hatch incubation conditions Control the temperature at 38.2°C, hatch for 10 days, the relative humidity is 60% on the 1st to 7th day, and 70% on the 8th to 10th day; 2018.11.02 by 10 5.0 EID 50 The inoculation dose per piece is used to inoculate chicken infectious bronchitis virus H52 strain in the allantoic cavity inoculation method; the post-hatch hatching conditions are controlled at a temperature of 37.8°C and a relative humidity of 70%; 24 hours after inoculation, the dead embryos are removed and the virus is inoculated. After 36 hours, the chicken embryos were placed in a 2-8°C refrigerator, and the virus allantoic fluid was harvested after about 12 hours of refrigeration. The virus content of this batch of chicken infectious bronchitis virus anti...

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Abstract

The invention belongs to the technical field of vaccine antigens, and discloses a method for preparing infectious bronchitis viral antigen by adopting non-immune eggs. The method comprises the following steps that infectious bronchitis non-immune eggs are adopted, pre-incubation conditions are controlled for pre-incubation; toxic strains of infectious bronchitis virus are inoculated on a chick embryo, and the inoculation dose is 105.0EID50/piece; after inoculation, post-incubation conditions are controlled for post-incubation, inspecting is still conducted after inoculation for 24 h, a dead embryo is discarded, after inoculation for 36 h, the chick embryo is placed into a cold storage chamber at 2-8 DEG C, and a virus allantoic fluid is obtained after 12 hours of cold storage; the virus titer of the obtained virus allantoic fluid is detected, and the virus allantoic fluid is concentrated and purified to obtain the infectious bronchitis viral antigen. According to the method, SPF eggs are replaced by the non-immune eggs, meanwhile the process conditions of the infectious bronchitis virus on the propagation of the non-immune eggs are determined, the quality of vaccine is ensured, thecost of vaccine research and production is also lowered, the huge reference value can be provided for vaccine manufacturing enterprises, and application prospects are broad.

Description

technical field [0001] The invention belongs to the technical field of vaccine antigens, and relates to a method for preparing chicken infectious bronchitis virus antigens by using non-immune eggs. Background technique [0002] Chicken infectious bronchitis (Infectious bronchitis, IB) is an acute, highly contagious infectious disease of chickens caused by chicken infectious bronchitis virus (Infectious bronchitis virus, IBV). Lesions of multiple tissues and organs such as the tract, glandular stomach, etc., mainly cause disease in chickens under 10 weeks old, with tracheal rales, coughing, and sneezing as the main clinical features. ", a serious threat to the development of my country's chicken industry. Due to the complex serotypes of chicken infectious bronchitis, there is no or only partial cross-immune protection between them, which brings a lot of inconvenience to the prevention and control of chicken infectious bronchitis disease. [0003] Vaccine immunization is an ef...

Claims

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Application Information

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IPC IPC(8): A61K39/215A61P31/14
CPCA61K39/12A61P31/14A61K2039/552C12N2770/20034
Inventor 李厚伟姚奋卯王彦辉彭丽孟蕾张盼盼孟祥李冰张宁姬星宇
Owner 商丘美兰生物工程有限公司
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