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为了解广东地区猪流感病毒(SIV)的流行和变异情况,本研究于2013年10月至2014年1月从广东省4个不同地区采集猪鼻拭子和肺脏病料共203份,对样品处理之后进行鸡胚分离和RT-PCR检测,从中分离得到5株SIV,对其进行病毒纯化、全基因组测序和遗传进化分析。结果表明,5株SIV分离毒株其中3株为H1N1亚型,2株为H1N2。分离毒株HA裂解位点位于PSIQSR↓GL,具有典型的低致病性流感病毒特征。HA基因序列比对结果显示,5株SIV分离毒株与A/Jiangsu/ALS1/2011(H1N1)株同源性最高,核苷酸相似性为97%~99%。遗传进化分析结果显示,5株分离毒株的PB2、PB1、PA和NP基因片段属于PDM/09分支,M基因片段以及外部基因片段HA属于欧亚类禽分支,NS基因片段属于北美三元重组分支,3株H1N1亚型的NA基因属于欧亚类禽分支,2株H1N2亚型的NA基因属于人季节性流感分支。抗原位点分析结果表明,分离毒株的抗原位点基本保守,但YJ28分离株HA1蛋白上发现3个氨基酸突变,分别是L69S、S137P、E222G。本研究通过对广东省不同地区分离到的SIV进行鉴定分析,掌握SIV的变异趋势,为猪流感的防控提供切实有效的理论依据。
In order to understand the prevalence and variation of swine influenza virus (SIV) in Guangdong, a total of 203 swine nasal swabs and lung disease samples were collected from 4 different areas of Guangdong Province from October 2013 to January 2014, After that, the chicken embryos were separated and detected by RT-PCR. Five SIV strains were isolated and purified. The whole genome was sequenced and analyzed for genetic evolution. The results showed that three isolates from five SIV strains were H1N1 subtype and two strains were H1N2. Isolates HA cleavage site is located in PSIQSR ↓ GL, with the typical characteristics of low pathogenic influenza virus. Sequence analysis of HA gene showed that the five strains isolated from SIV had the highest homology with A / Jiangsu / ALS1 / 2011 (H1N1), and the nucleotide similarity was 97% -99%. The results of genetic evolution analysis showed that the PB2, PB1, PA and NP gene fragments of the five isolates belonged to the PDM / 09 branch, the M gene fragment and the outer gene fragment HA belonged to the Eurasian avian poultry branch, and the NS gene fragment belonged to the North American ternary recombination The NA genes of three H1N1 subtypes belong to the Eurasian poultry branch. The two NA genes of the H1N2 subtype belong to the human seasonal influenza branch. Antigenic site analysis showed that the antigenic sites of the isolated strains were basically conserved, but three amino acid mutations were found on the HA1 protein of YJ28 isolate, namely L69S, S137P and E222G. In this study, through the identification of SIV isolated in different areas of Guangdong Province to master the trend of variation of SIV, to provide an effective and effective theoretical basis for the prevention and control of swine influenza.