Pig replication and respiration syndrome virus attenuated vaccine strain and uses thereof
A technology for respiratory syndrome and attenuated vaccines, applied in the field of porcine reproductive and respiratory syndrome virus attenuated vaccine strains and attenuated vaccine strains, can solve the problems of poor vaccine pertinence, unsatisfactory immune effect, antigenic variation, etc., and achieve good safety Effect
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[0021] 1.1 The pigs used in the test were purchased from PRRS seronegative pig farms in non-epidemic areas.
[0022] 1.2 Virus cell culture and cloning
[0023] 1.2.1 Cell culture Marc-145 cell line was cultured with DMEM cell culture medium containing 4% inactivated calf serum, 100 U / ml penicillin, 80 μg / ml streptomycin, and NaHCO 3 Adjust the pH value to about 7.2, and filter to sterilize. Use this cell culture solution to resuscitate or subculture the cell line Marc-145 into a sterilized small flat bottle with a capacity of 100ml, and use it to inoculate the virus after it grows into a good cell monolayer.
[0024] 1.2.2 Cloning of the virus The PRRSV CH-1a strain isolated from China was passaged on Marc-145 cells, and virus clones were carried out at the 30th, 50th, 70th, 90th, 110th, and 130th generations, except for the 30th generation A 2mm plaque was selected for seeding clones, and a 1mm plaque was selected for subsequent clones. The 30th and 50th generation seed p...
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