摘要
从广东省不同猪场分离到4株H3N2亚型猪源流感病毒A／Swine／Guangdong／01／2004、A／Swine／Guang-dong／02／2004、A／Swine／Guangdong／03／2004、A／Swine／Guangdong／04／2004。根据GenBank公布的H3N2亚型猪源流感病毒的HA基因序列，设计1对引物，运用RT-PCR方法扩增四株病毒的HA基因，并进行测序和分析。同源性分析和遗传进化分析表明本实验的4株H3N2亚型SIVHA基因核苷酸序列同源性为99.8％-99.9％，在遗传进化树中均位于同一分支上。与参考毒株的比较分析表明，4个毒株与WHO推荐的2001-2004年北半球H3N2亚型流感疫苗株A／Moscow／10／99HA基因的核苷酸序列同源性最高为99.4％-99.5％，4个毒株与A／Moscow／10／99HA基因在遗传进化树中位于同一个小分支上。氨基酸序列比较发现，4个毒株HA基因裂解位点处的氨基酸序列均为PEKQTR↓G，4个毒株推导的氨基酸序列中均有11个糖基化位点，4个毒株HA蛋白226位受体结合位点（RBS）处氨基酸均为异亮氨酸（I）。4个毒株HA基因的氨基酸序列、受体结合位点以及糖基化位点均与A／Moscow／10／99相应的氨基酸序列一致。本试验的4株H3N2亚型猪源流感病毒的HA基因属于以A／Moscow／10／99为代表的近代类人H3N2亚型流感病毒，在一定程度上揭示了广东省H3N2亚型猪流感病毒HA基因进化与流行情况。
Four strains of H3N2 subtype swine influenza viruses, named as A/Swine/Guangdong/01/2004, A/ Swine/Guangdong/02/2004, A/Swine/Guangdong/03/2004, A/Swine/Guangdong/04/2004, were isolated from pigs with influenza-like signs in Guangdong province in 2004. Reverse transcription-polymerase chain reaction （RT-PCR） was used to amplify the HA gene segments for sequencing analysis. As demonstrated by homology and phylogenic a- nalysis,it was found that higher homologues in the corresponding nucleotide sequence and closed relationship existed in these strains of SIV and also with the recommented virus A/Moscow/10/99. As demonstrated by comparison of the amino acid sequences,it was found that the cleavage site between HA1 and HA2 was PEKQTR ↓ G and the 226 position amino acid of HA was I . The amino acid of HA contained 11 glycosylation sites and ten of them existed in the HA1 gene,one in HA2 gene. It is concluded that the HA gene of these four H3N2 SIV strains isolated from Guangdong province may be from the same source with HA gene of influenza virus A/Moscow/10/99, belonging to recent human H3N2 subtype influenza virus.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2009年第11期1424-1428,共5页
Chinese Journal of Veterinary Science
基金
广东省科技攻关项目(2006B20801002)
