Attenuated method for fowl infectious bronchitis viruses
A bronchitis and infectious technology, which is applied in the attenuation field of chicken infectious bronchitis virus, can solve the problems of slow attenuation speed, achieve the effects of reducing loss, saving passage time, and shortening attenuation time
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Embodiment 1
[0025] Embodiment 1 continuous passage weakens
[0026] A method for attenuating avian infectious bronchitis virus. After diluting the virus strain YX10 by 10 times by 100 times, inoculate 3 SPF chicken embryos that have been incubated at the corresponding culture temperature for 10 hours, and inoculate each embryo with 103.5-4EID50 / 0.2 mL, after culturing for a certain period of time, put the chicken embryos at -20°C for half an hour, collect the allantoic fluid aseptically, and carry out the next generation and sub-passage; the culture temperature from the first generation to the tenth generation is 37°C, and the culture time is 48 hours ;The culture temperature of the 11th generation to the 14th generation is reduced by 1°C for each generation, that is, the culture temperature of the 11th generation is 36°C, the culture temperature of the 12th generation is 35°C, the culture temperature of the 13th generation is 34°C, and the culture temperature of the 14th generation is 33...
Embodiment 2
[0027] Embodiment 2 continuous passage weakens
[0028] A method for attenuating avian infectious bronchitis virus. After diluting the virus strain YX10 by 10 times to 100 times, inoculate 3 SPF chicken embryos that have been incubated at the corresponding culture temperature for 12 hours, and each embryo is inoculated with 10 3.5-4 EID 50 / 0.2mL, after culturing for a certain period of time, put the chicken embryo at -20°C for half an hour, collect the allantoic fluid aseptically, and carry out the next generation and subculture; 48 hours; from the 11th generation to the 14th generation, the culture temperature is reduced by 1°C for each generation, and the culture time is 48 hours; from the 15th generation onwards, the culture temperature is 32°C, and the culture time from the 15th generation to the 60th generation is 40 hours; From generation 61 onwards, the culture time is 32 hours.
Embodiment 3
[0029] Embodiment 3 continuous passage weakens
[0030] A method for attenuating avian infectious bronchitis virus. After diluting the virus strain YX10 by 10 times to 100 times, inoculate 3 SPF chicken embryos that have been incubated at the corresponding culture temperature for 8 hours, and each embryo is inoculated with 10 3.5-4 EID50 / 0.2mL, after culturing for a certain period of time, put the chicken embryo at -20°C for half an hour, collect the allantoic fluid aseptically, and carry out the next generation and subculture; 48 hours; from the 11th generation to the 14th generation, the culture temperature is reduced by 1°C for each generation, and the culture time is 48 hours; from the 15th generation onwards, the culture temperature is 32°C, and the culture time from the 15th generation to the 60th generation is 44 hours; From generation 61 onwards, the culture time is 36 hours.
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