H_3N_2亚型猪源流感病毒NS基因的克隆与序列分析

Cloning and Sequence Analysis of NS gene of H_3N_2 Swine Influenza Virus

根据GenBank上已发表的H3N2亚型的猪流感病毒NS基因序列,设计合成一对引物,对四个猪流感病毒广东株提取RNA,运用RT-PCR方法获得了4株H3N2亚型猪源流感病毒广东分离株A/Swine/Guangdong/01/04(H3N2)、A/Swine/Guangdong/02/04(H3N2)、A/Swine/Guangdong/03/04(H3N2)、A/Swine/Guangdong/04/04(H3N2)的NS基因序列,并对所得序列进行比较分析。序列分析表明,四个毒株NS基因的核苷酸序列与参考毒株的NS基因相似性高达90%以上,说明近年来广东的H3N2亚型猪流感病毒NS基因变异较小。遗传进化关系表明本实验四个毒株的NS基因和香港2000-2002年间的猪流感病毒的NS基因与1998-2002年间美洲流行的人流感病毒的NS基因在遗传进化树中位于同一个分支,这说明它们可能有着相近的亲缘关系。

A pair of primers were designed according to the NS gene sequences of H3N2 subtype swine influenza virus（SIV） in NCBI. The NS genes of four H3N2 SIV strains isolated from Guangdong province were successfully amplified by RT-PCR. Sequence analysis indicated that the four NS nucleotide sequences had more than 90% identity with the reference sequences. It showed that the NS gene sequences of H3N2 subtype SIV had little variation in recent years. Phylogenic analysis showed that the NS genes of these four SIV strains had close relationship with the NS genes of H3N2 subtype SIV swains isolated in Hongkong from year 2000 to 2002 and strains isolated in America from year 1998 to 2002.