Objective To apply the next generation sequencing(NGS)technology to the quality control of Sabin strain inactivated poliomyelitis vaccines(sIPVs)(Vero cells)during the production,in order to ensure the safety and inter-batch consistency of the vaccines.Methods The poliovirus(PV)working seed lot of Sabin strain Ⅰ,Sabin strain Ⅱ,Pfizer strain Ⅲ were inoculated on the monolayer Vero cells cultured with microvectors in a bioreactor at 33 ℃ for 3-4 days to prepare vaccinal virus liquid,three batches for each type.RNA was extracted from all types of working seeds and vaccinal virus liquid,and the genome mutation frequency,neurovirulence determinants,mutation hotspots and inter-batch consistency were ana-lyzed by NGS.Results Sabin strain Ⅱ and Pfizer strain Ⅲ were more vulnerable to mutating than Sabin strain Ⅰ after the culture of three types PV of working seed lot,and the mutation frequency of neurovirulence determinants increased signifi-cantly in non-coding region(t=3.21-5.83,each P＜0.05),while changed little in coding region(t=1.29-2.34,each P＞0.05)or decreased significantly(t=6.01-9.62,each P＜0.05).There was a mutation hotspot at nt869 of Sabin strain Ⅱ and nt2 493 of Pfizer strain Ⅲ in the manner of C→T respectively,and nt2 493 was not only a neurovirulence determinant,but also a mutation hotspot.Three batches of vaccinal virus liquid of three types all showed good inter-batch consistency,and the inter-batch fitted R2 of mutation frequency of nucleic acid sites in genome VP1 region ranged from 0.947 to 0.995.Conclusion The NGS technique can efficiently detect the mutations of PV genome in the production of sIPV,which is stable and reliable,and can be used for the quality control of the vaccine.
Next generation sequencing(NGS)Sabin strain inactivated poliomyelitis vaccine(sIPV)Quality controlMutation frequency
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