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To screen primary chicken embryo cells adapted strain of live attenuated yellow fever vaccine 17D-204，prepare virus seed banks and perform release tests.
The virus seed of live attenuated yellow fever vaccine 17D-204 was adapted and domesticated on primary chicken embryo cells，and the chicken embryo cells adapted strain was obtained. Establish virus seed banks of live attenuated yellow fever vaccine (primary chicken embryo cells)，and quality control was performed.
The optimum culture temperature of live attenuated yellow fever vaccine 17D-204 in primary chicken embryo cells was 37 ℃，and the optimum MOI(multiplicity of infection) was 0.0048. Three-level seed banks of primary seed bank，master seed bank，working seed bank of live attenuated yellow fever vaccine 17D-204 (primary chicken embryo cells) [17D-204(CEC)] were established. The verification of virus titer，identification test，sterility test，mycoplasma，exogenous virus factor test，Mycobacterium test and other items were in line with the provisions of volume Ⅲ of the Chinese Pharmacopoeia. High throughput sequencing showed that there was no gene mutation and no internal and external virus factor pollution in the gene sequence of the virus seed banks.
The primary chicken embryo cell adapted strain of live attenuated yellow fever vaccine 17D-204(CEC) can be screened out to establish the virus seed banks，with good passage stability and genetic stability. This study laid a firm foundation for the regeneration of live attenuated yellow fever vaccine from chicken yolk sac to chicken embryo cells.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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