无细胞百日咳疫苗生产用菌株CS的5种保护性抗原基因序列分析

Gene Sequencing of Five Kinds of Protective Antigens of CS Strain for Production of Acellular Pertussis Vaccine

目的对我国无细胞百日咳疫苗生产用菌株CS的5种保护性抗原的基因序列进行分析。方法分别用PCR扩增CS菌株的PT、FHA、PRN、FIM2和FIM3基因,克隆至pGEM-TEasy vector中,进行序列测定及分析,并确定其基因型。结果获得了CS菌株PT、FHA、PRN、FIM2和FIM35段基因克隆,其核苷酸和氨基酸序列与GenBank中收录的相应百日咳杆菌的同源性均在90%以上,种系进化树分析结果与鲍特菌属进化结果一致。CS的基因型为:PTS1属于ptxS1B型,PTS3属于ptxS3A型,FHA属于FHA1型,PRN属于PRN1型,FIM2属于FIM2-1型,FIM3属于FIM3A型。结论本研究为更好地对我国无细胞百日咳疫苗进行质量控制和百日咳杆菌的基因漂移研究奠定了基础。

Objective To analyze the gene sequences of five kinds of protective antigens of CS strain for production of acellular pertnssis vaccine in China. Methods The PT, FHA, PRN, FIM2 and FIM3 genes of CS strain were amplified by PCR and cloned into pGEM-T Easy vector for gcne sequencing, based on which the genotypes were determined. Results The PT, FHA, PRN, FIM2 and FIM3 gene clones were obtained, of which the homologies of nucleotide and amino acid sequences were more than 90% to, and the results of analysis by phylogenetic tree were consistent with those of Bordetella pertussis reported in GenBank. The PT S1, PTS3, FHA, PRN, FIM2 and FIM3 of CS strain were of genotypes of ptXS1B, ptxS3A, FHA1, PRN1, FIM2-1 and FIM3A respectively. Conclusion The gene sequences of five kinds of protective antigens of CS strain were analyzed, which laid a foundation of further quality control of acellular pertussis vaccine in China and study on the gene drift of Bordetella pertussis.