河北省甲型H_1N_1流行性感冒病毒血凝素基因特征研究

Genetic Characteristics of the Haemagglutinin Genes of Pandemic(H_1N_1) 2009 Viruses in Hebei Province

目的掌握河北省甲型H1N1流行性感冒(甲流)病毒的流行情况;对分离到的甲流病毒株血凝素(Hemagglutinin,HA)基因序列进行测定比较,分析HA重链区(HA1)第222位氨基酸位点的突变情况。此位点编码的天冬氨酸(Aspartic Acid,N)突变为甘氨酸(Glycine,G)(D222G),可能会使病毒更容易结合下呼吸道受体,为流感大流行的预防控制提供参考。方法采集流感样病例的鼻咽拭子,用狗肾传代细胞分离流感病毒,经血凝抑制试验鉴定分型。选取不同时间、地域的10株甲流病毒进行HA基因全序列分析,167株病毒测HA区379～1204片段,用DNA(脱氧核糖核酸,Deoxyribonucleic Acid)Star软件比较分析同源性、变异趋势。结果 10株甲流病毒毒株HA基因与疫苗株A(甲)/California(加利福尼亚)/07/2009相比高度同源,核苷酸同源性为98.9%～99.5%,氨基酸同源性为98.4%～99.3%,有3个位点氨基酸发生同样的替换(脯氨酸,Proline)100S(丝氨酸,Serine),S220T(苏氨酸,Threonine),E(谷氨酸,Glutamic Acid)391K(赖氨酸,Lysine)。从采集于2010年12月标本分离的2株甲流病毒A/河北桥东/SWL(Swine-like)178/2010和A/河北裕华/SWL1261/2010,在202、214、468三个氨基酸位点与其他毒株不同。所有167株甲流病毒的HA1第222位氨基酸均未发生D222G突变。结论河北省甲流病毒HA基因与疫苗株A/California/07/2009高度同源,与疫苗株及各毒株之间核苷酸同源性均≥98.9%,尚未发现D222G变异。

Objective To study the pandemic （HIN1） 2009 viruses activity and the characteristics of hemagglutinin （HA） genes of pandemic （HINi） 2009 viruses isolated in Hebei province. To analyze whether the isolated viruses represented such mutation as the amino acid change from aspartic acid （D） to glycine （G） at residue 222 might giving a rise to the combination of virus to lower respiratory tract receptor and to provide the scientific basis for influenza pandemic. Methods The pharynx swabs specimens were inoculated into madin-darby canine kidney for influenza virus type and subtype by he cell flasks for influenza virus. The isolates were identified magglutination inhibition assay.10 Pandemic （H1N1） 2009 strains were chosen by different time to sequence HA gene.167 strains were detected the segment from 379 to 1204 of HA gene. The sequences of deoxyribonucleic acid and amino acid were compared with that of the representative strains by DNA （Deoxyribonucleic Acid）Star software. Results The HA gene of ten Pandemic （H1N1） 2009 strains were quite closely related to that of vaccine strain A/ California/07/2009, with amino acid homology 98.4%-99.3 % and the nucleotide homology 97.5 %-97.8 % with each other. Three amino acids were substituted [- P （ Proline ） 100S （Serine）, S220 T （Threonine） ,E （ Glutamic Acid） 391K （Lysine） ]. TheA/Hebeiqiaodong/SWL （ swine-like ） 178/2010 and A/Hebeiyuhua/ SWL1261/2010 isolated in December 2010 were different from other strains at the amino acide 202,214 and 468. There was no mutation at the position D222G of the HA1 gene among all the 167 pandemic （H1N1）2009 strains in Hebei province. Conclusion The HA gene of pandemic viruses from Hebei are quite closely related to that of A/California/07/2009 vaccine virus, with ≥ 98.9% homology. There is no mutation at the position D222G of the HA1 gene among pandemic viruses in Hebei province.