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Preparation method of newcastle disease HI antigen

A technology for Newcastle disease and antigens, applied in biochemical equipment and methods, virus antigen components, antiviral agents, etc., can solve the problems of lack of safety, stability, and easy storage of antigens in diagnostic antigens, and achieve stable and convenient HA hemagglutination activity Effect of saving and reducing workload

Active Publication Date: 2012-02-15
北京盛华四合生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to disclose a method for preparing Newcastle disease HI antigen to overcome the problem of lack of safe, stable and easy-to-storage antigens in the existing Newcastle disease HI diagnostic antigens

Method used

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  • Preparation method of newcastle disease HI antigen
  • Preparation method of newcastle disease HI antigen
  • Preparation method of newcastle disease HI antigen

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] PEG4000 from Beijing Xinjingke Biotechnology Co., Ltd. was used as the main component of the stabilizer. The preparation steps of HI stable antigen are as follows:

[0032] (1) SPF chicken embryos (provided by Beijing Meria Weitong Experimental Technology Co., Ltd.) were inoculated with poison

[0033] Under sterile conditions, the Newcastle disease La Sota strain virus was diluted to contain 10 6 EID5 0 / 0.1ml, inoculated with well-developed SPF chicken embryos aged 9-11 days, 0.1ml / piece, sealed with wax after inoculation.

[0034] (2) Culture and Embryo

[0035] After inoculation, the SPF chicken embryos were incubated at 37°C, and the eggs were illuminated once every 24 hours, and the dead embryos were discarded. After that, the eggs were illuminated once every 6 hours, and the dead chicken embryos were taken out in time and placed at 4°C. ℃ overnight.

[0036] (3) Harvest and HA detection

[0037] Harvest chicken embryo allantoic fluid under aseptic environme...

Embodiment 2

[0051] Using PEG8000 from Beijing Xinjingke Biotechnology Co., Ltd. as the main component of the stabilizer, the preparation process steps of HI stabilized antigen are as follows:

[0052] (1) Inoculation of SPF chicken embryos

[0053] Under sterile conditions, the Newcastle disease La Sota strain virus was diluted to contain 10 6 EID 50 / 0.1ml, inoculated with well-developed SPF chicken embryos at the age of 9-11 days, 0.1ml / piece, sealed with wax.

[0054] (2) Culture and Embryo

[0055] After inoculation, the SPF chicken embryos were incubated at 37°C, and the eggs were illuminated once every 24 hours, and the dead chicken embryos were discarded. After that, the eggs were illuminated once every 6 hours, and the dead chicken embryos were taken out in time and placed at 8°C until 96 hours. ℃ overnight.

[0056] (3) Harvest and HA detection

[0057] Harvest chicken embryo allantoic fluid under aseptic environment, and do sterility test at the same time. Detection of HA ...

Embodiment 3

[0071] Inactivation test: After diluting the inactivated allantoic fluid in Example 1 and Example 2 by 1:10000 times, inoculate 11-day-old SPF chicken embryos through the allantoic cavity, 0.1 mL per embryo, and continue at 37 ° C after inoculation After hatching for 5 days, observe the development of the chicken embryo and detect whether the allantoic fluid of the chicken embryo has hemagglutination activity. The harvested chicken embryo allantoic fluid was blindly passed on SPF chicken embryos for three generations, and the chicken embryo allantoic fluid had no hemagglutination activity. It shows that the antigen prepared by the invention is completely inactivated, safe and non-toxic.

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Abstract

The invention provides a preparation method of a newcastle disease HI antigen. The method comprises the following technical steps: (1) inoculating newcastle disease virus into a chick embryo, then harvesting allantoic fluid of the chick embryo and adding an inactivator, namely beta-propiolactone according to 0.25 per mill-0.5 per mill by volume of the allantoic fluid; (2) concentrating the inactivated allantoic fluid to 1 / 10-1 / 20 of the original volume and adding a stabilizer; (3) further using PBS (phosphate buffer saline) buffer solution in the volume which is 1 / 5-1 / 10 of that of the allantoic fluid before concentration for diluting the antigen; and (4) performing ultrasonic degradation, adding glycerol which accounts for 20%-30% of the final volume, oscillating and uniformly mixing so as to get the antigen which can be used. The prepared antigen has no risk of dispersing toxin during the use and the storage, and can reduce the damage to an environment and a human body due to the use of the live virus antigen, the product has good stability, the hematic coagulating rate of the product is unchanged by being preserved for above 24 months at the temperature of 2-8 DEG C, and the newcastle disease HI antigen can be used for detecting newcastle disease HI.

Description

technical field [0001] The invention relates to a method for preparing Newcastle disease HI antigen, belonging to the field of biotechnology. Background technique [0002] Newcastle disease (ND), also known as Asian chicken plague or pseudo chicken plague, is an acute, highly contagious and fatal poultry infectious disease caused by Newcastle disease virus of the genus Paramyxovirus. The disease was first discovered in Indonesia in 1926, and in Newtown, England in the same year. The disease was first reported in my country in 1935. It was considered to be bird flu at that time, and it was confirmed to be Newcastle disease in 1948. There have been three pandemics of the disease in the world. By the end of the 1980s, new changes occurred in the disease, which mostly occurred in chicks with maternal antibodies and immunized chickens, and the clinical symptoms were not typical; in immunized chickens In laying hens, there are non-acute, non-acute, low-lethal, and unobvious sympt...

Claims

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Application Information

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IPC IPC(8): C12N7/00A61K39/17A61P31/14
Inventor 王世敏王文泉张洪郑杰张红张发明杨国良孙丰廷陈玲李静
Owner 北京盛华四合生物科技有限公司
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