Preparation method of triple vaccine for treating avian H3N8 influenza virus
By developing a trivalent inactivated vaccine, the problem of requiring multiple injections with existing vaccines has been solved. This vaccine provides simultaneous protection against H3N8 influenza virus, Newcastle disease, and avian adenovirus, reducing stress and costs, and improving the efficiency of healthy poultry growth.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-11-06
- Publication Date
- 2026-03-13
AI Technical Summary
Existing poultry vaccines can only protect against a single virus, requiring multiple injections, which leads to stress and a heavy workload for farmers, and cannot effectively prevent multiple diseases.
A trivalent vaccine containing H3N8 influenza virus, Newcastle disease virus, and avian adenovirus was prepared by virus culture, inactivation, mixing of oil and water phases, and emulsification to form an inactivated vaccine, enabling prevention of three diseases with a single vaccination.
Improve disease prevention efficiency, reduce stress response, lower costs, ensure comprehensive immune protection in a short period of time, reduce the number of vaccinations, and promote healthy growth of poultry.
Abstract
Description
Technical Field
[0001] This invention belongs to the field of poultry vaccine preparation technology, specifically relating to a method for preparing a trivalent vaccine for treating avian H3N8 influenza virus. Background Technology
[0002] H3N8 avian influenza virus belongs to the type A influenza virus and is highly variable and transmissible. This virus primarily infects poultry, including chickens, ducks, geese, and other domestic birds, and can also infect some wild birds. H3N8 avian influenza virus spreads rapidly and over a wide area; once an outbreak occurs, it often causes huge economic losses to the poultry industry. Newcastle disease (ND), commonly known as fowl plague, is a highly contagious, acute, and deadly infectious disease of chickens caused by the Newcastle disease virus. It has a high morbidity and mortality rate and occurs worldwide. Once an outbreak occurs, it can have a devastating impact on the poultry farming industry. Avian adenovirus can cause inclusion body hepatitis and pericardial effusion syndrome in poultry. Clinically, it is characterized by liver degeneration and necrosis, and a pale yellow, clear effusion in the pericardial cavity. It poses a significant threat to the development of the poultry industry, with a mortality rate that can reach 80%.
[0003] Most current poultry vaccines can only protect against a single virus. Therefore, poultry farmers need to vaccinate their birds multiple times during the breeding process, which places a heavy burden on both farmers and poultry. Multiple vaccinations can also easily cause stress reactions in poultry, affecting their growth, health, and growth rate. Summary of the Invention
[0004] To address the problems mentioned in the background section, this invention provides a method for preparing a trivalent vaccine for treating avian H3N8 influenza virus, which features comprehensive preventative effect and reduced stress response.
[0005] To achieve the above objectives, the present invention provides the following technical solution: a method for preparing a trivalent vaccine for treating avian H3N8 influenza virus, comprising the following steps: Step 1: Virus culture. H3N8 influenza virus and Newcastle disease virus were inoculated into chicken embryos, respectively. After culture, the chicken embryo fluid containing H3N8 influenza virus and Newcastle disease virus was harvested, and then ultrafiltration was performed to concentrate the H3N8 influenza virus stock solution and Newcastle disease virus stock solution. Avian adenovirus was inoculated into LMH cells, cultured, and then separated and concentrated by ultrafiltration to obtain avian adenovirus stock solution.
[0006] Step 2: Virus inactivation. Formaldehyde solution is added to the obtained virus stock solution for inactivation. After inactivation, the virus stock solution is stored at low temperature.
[0007] Step 3: Oil phase preparation. Take 94 parts of white oil and 1 part of aluminum stearate, mix them evenly and heat them. Then add 5 parts of Span 80, heat and mix evenly, and sterilize.
[0008] Step 4: Aqueous phase preparation. Mix the extinguished H3N8 avian influenza virus stock solution, Newcastle disease virus stock solution, and avian adenovirus stock solution. Take 96 portions of the mixed antigen solution and 4 portions of sterilized Tween-80, and mix thoroughly.
[0009] Step 5: Emulsification preparation. The prepared oil phase solution is mixed with the aqueous phase solution and emulsified to obtain the trivalent inactivated vaccine.
[0010] Preferably, the virus inactivation uses a formaldehyde concentration of 2.5% and is performed at 35°C for 20 hours.
[0011] Preferably, after the virus stock solution is inactivated, it needs to undergo sterilization and inactivation testing. The inactivated virus stock solution should be cultured and should show no sterile growth.
[0012] Preferably, the oil phase preparation conditions are as follows: white oil and aluminum stearate are mixed evenly in an oil phase preparation tank and heated to 75°C, then Span 80 is added, and the temperature is maintained at 120°C for 40 minutes, and then cooled by purging with cold water.
[0013] Preferably, in the aqueous phase preparation process, sterilized Tween-80 is added to the mixed antigen solution and stirred continuously until Tween-80 is completely dissolved.
[0014] Preferably, the oil phase and the water phase are mixed and emulsified in a volume ratio of 2:1.
[0015] Compared with the prior art, the beneficial effects of the present invention are: 1. This invention, the H3N8 influenza virus trivalent vaccine, can improve the efficiency of poultry disease prevention, reduce disease prevention costs, reduce stress response, and prevent multiple diseases. A single vaccination can prevent three poultry diseases simultaneously, ensuring that poultry can obtain comprehensive immune protection in a short period of time, reducing the number of vaccinations, helping to reduce the stress response of poultry caused by vaccination, which is conducive to the healthy growth of poultry, and at the same time reducing the workload of farmers. Detailed Implementation
[0016] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.
[0017] Example 1 This invention provides the following technical solution: a method for preparing a trivalent vaccine for treating avian H3N8 influenza virus, comprising the following steps: Step 1: Virus culture. H3N8 influenza virus and Newcastle disease virus were inoculated into chicken embryos, respectively. After culture, the chicken embryo fluid containing H3N8 influenza virus and Newcastle disease virus was harvested, and then ultrafiltration was performed to concentrate the H3N8 influenza virus stock solution and Newcastle disease virus stock solution. Avian adenovirus was inoculated into LMH cells, cultured, and then separated and concentrated by ultrafiltration to obtain avian adenovirus stock solution.
[0018] Step 2: Virus inactivation. Formaldehyde solution is added to the obtained virus stock solution for inactivation. After inactivation, the virus stock solution is stored at low temperature.
[0019] Step 3: Oil phase preparation. Take 94 parts of white oil and 1 part of aluminum stearate, mix them evenly and heat them. Then add 5 parts of Span 80, heat and mix evenly, and sterilize.
[0020] Step 4: Aqueous phase preparation. Mix the extinguished H3N8 avian influenza virus stock solution, Newcastle disease virus stock solution, and avian adenovirus stock solution. Take 96 portions of the mixed antigen solution and 4 portions of sterilized Tween-80, and mix thoroughly.
[0021] Step 5: Emulsification preparation. The prepared oil phase solution is mixed with the aqueous phase solution and emulsified to obtain the trivalent inactivated vaccine.
[0022] Specifically, the formaldehyde concentration used for virus inactivation was 2.5%, and the virus was inactivated at 35°C for 20 hours.
[0023] Specifically, after the virus stock solution is inactivated, it needs to be tested for sterilization and inactivation. The inactivated virus stock solution should be cultured and should grow sterilely.
[0024] Specifically, the oil phase preparation conditions are as follows: white oil and aluminum stearate are mixed evenly in an oil phase preparation tank and heated to 75°C. Then, Span 80 is added, and the temperature is maintained at 120°C for 40 minutes. Finally, cold water is circulated to cool the mixture down.
[0025] Specifically, in the aqueous phase preparation process, sterilized Tween-80 is added to the mixed antigen solution and stirred continuously until Tween-80 is completely dissolved.
[0026] Specifically, the oil phase and the water phase are mixed and emulsified in a volume ratio of 2:1.
[0027] Finally, it should be noted that the above descriptions are merely preferred embodiments of the present invention and are not intended to limit the present invention. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions described in the foregoing embodiments or make equivalent substitutions for some of the technical features. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.
Claims
1. A method for preparing a trivalent vaccine for treating avian H3N8 influenza virus, characterized in that, Includes the following steps: Step 1: Virus culture. H3N8 influenza virus and Newcastle disease virus were inoculated into chicken embryos, and after culture, chicken embryo fluid containing H3N8 influenza virus and Newcastle disease virus was harvested. Then, ultrafiltration was performed to concentrate the H3N8 influenza virus stock solution and Newcastle disease virus stock solution. Avian adenovirus was inoculated into LMH cells, cultured, and then separated and concentrated by ultrafiltration to obtain avian adenovirus stock solution. Step 2: Virus inactivation. Formaldehyde solution is added to the obtained virus stock solution for inactivation. After inactivation, the virus stock solution is stored at low temperature. Step 3: Oil phase preparation: Take 94 parts of white oil and 1 part of aluminum stearate, mix them evenly and heat them, then add 5 parts of Span 80, heat and mix them evenly, and sterilize them. Step 4: Aqueous phase preparation. Mix the extinguished H3N8 avian influenza virus stock solution, Newcastle disease virus stock solution, and avian adenovirus stock solution. Take 96 parts of the mixed antigen solution and 4 parts of the sterilized Tween-80 solution and mix thoroughly. Step 5: Emulsification preparation. The prepared oil phase solution is mixed with the aqueous phase solution and emulsified to obtain the trivalent inactivated vaccine.
2. The method for preparing a trivalent vaccine for treating avian H3N8 influenza virus according to claim 1, characterized in that: The virus was inactivated using a formaldehyde concentration of 2.5% at 35°C for 20 hours.
3. The method for preparing a trivalent vaccine for treating avian H3N8 influenza virus according to claim 1, characterized in that: After the virus stock solution is inactivated, it needs to be tested for sterilization and inactivation. The inactivated virus stock solution should be cultured and should grow sterilely.
4. The method for preparing a trivalent vaccine for treating avian H3N8 influenza virus according to claim 1, characterized in that: The oil phase preparation conditions are as follows: white oil and aluminum stearate are mixed evenly in an oil phase preparation tank and heated to 75°C. Then, Span 80 is added, and the temperature is maintained at 120°C for 40 minutes. Then, cold water is circulated to cool it down.
5. The method for preparing a trivalent vaccine for treating avian H3N8 influenza virus according to claim 1, characterized in that: In the aqueous phase preparation process, sterilized Tween-80 is added to the mixed antigen solution and stirred continuously until Tween-80 is completely dissolved.
6. The method for preparing a trivalent vaccine for treating avian H3N8 influenza virus according to claim 1, characterized in that: The oil phase and water phase are mixed and emulsified at a volume ratio of 2:1.