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目的分析河北省2003～2008年流行期甲型流感病毒流行株对金刚烷胺的耐药情况,为流感的临床治疗用药提供参考。方法随机选取河北省流感监测网络中所分离的305株甲型流感病毒(甲1亚型76株,甲3亚型229株),提取病毒核酸,采用RT-PCR扩增流感病毒M2基因片段,纯化后进行核苷酸序列测定,用生物信息软件分析与耐药有关的氨基酸位点。结果 305株甲型流感病毒分离株中,250株M2蛋白跨膜区耐药位点的氨基酸被置换,对金刚烷胺耐药率为81.97%,2003～2008年5个流行期耐药率分别为90.32%、78.95%、29.63%、100%和100%。变异主要发生在M2蛋白跨膜区第31位氨基酸,244株发生了S31N置换,占97.60%,其次是在第27位有6株发生了V27A置换,占2.40%。结论河北省甲型流感病毒流行株对金刚烷胺的耐药率较高,2006～2008年流行期流行株耐药率达100%,提示临床治疗甲型流感不宜使用此类药物。
Objective To analyze the drug resistance of pandemic influenza A strains to amantadine in the epidemic period of 2003-2008 in Hebei Province and provide reference for the clinical treatment of influenza. Methods A total of 305 influenza A viruses isolated from influenza surveillance network in Hebei Province were randomly selected (76 strains of A 1, 229 strains of A 3) and the viral nucleic acids were extracted. The M2 gene fragment of influenza virus was amplified by RT-PCR, After purification, the nucleotide sequence was determined and bioinformatics software was used to analyze the drug resistance related amino acid sites. Results Among the 305 influenza A virus isolates, the amino acid substitutions of the resistance sites at 250 transmembrane regions of M2 protein were replaced, the resistance rate to amantadine was 81.97%, and the resistance rates at five epidemic phases from 2003 to 2008 were respectively 90.32%, 78.95%, 29.63%, 100% and 100%. The mutation mainly occurred in the amino acid at position 31 of the transmembrane region of M2 protein, S31N substitution occurred in 244 strains, accounting for 97.60%, followed by V27A substitution in 6 of 27 strains (2.40%). Conclusion The epidemic strains of influenza A in Hebei Province have a higher resistance rate to amantadine. The resistance rate of the epidemic strains in 2006 ~ 2008 reached 100%, suggesting that they should not be used in the clinical treatment of influenza A virus.