Świętoń Edyta, Śmietanka Krzysztof
Department of Poultry Diseases, National Veterinary Research Institute, 24-100 Pulawy, Poland.
J Vet Res. 2017 Dec 27;61(4):381-389. doi: 10.1515/jvetres-2017-0054. eCollection 2017 Dec.
The genomes of nine H5 subtypes of low pathogenic avian influenza virus (LPAIV) strains identified in wild birds in Poland between 2010 and 2015 were sequenced, and their phylogenetic relationship was determined.
AIV genome segments were amplified by RT-PCR and the PCR products were sequenced using Sanger method. Phylogenetic trees were generated in MEGA6 software and digital genotyping approach was used to visualise the relationship between analysed strains and other AIVs.
High genetic diversity was found in the analysed strains as multiple subgroups were identified in phylogenetic trees. In the HA tree, Polish strains clustered in two distinct subclades. High diversity was found for PB2, PB1, PA and NP, since 5-8 sublineages could be distinguished. Each strain had a different gene constellation, although relationship of as much as six out of eight gene segments was observed between two isolates. A relationship with poultry isolates was found for at least one segment of each Polish strain.
The genome configuration of tested strains indicates extensive reassortment, although the preference for specific gene constellation could be noticed. A significant relationship with isolates of poultry origin underlines the need for constant monitoring of the AIV gene pool circulating in the natural reservoir.
对2010年至2015年间在波兰野生鸟类中鉴定出的9种低致病性禽流感病毒(LPAIV)H5亚型毒株的基因组进行了测序,并确定了它们的系统发育关系。
通过逆转录聚合酶链反应(RT-PCR)扩增禽流感病毒基因组片段,并用桑格法对聚合酶链反应(PCR)产物进行测序。在MEGA6软件中生成系统发育树,并采用数字基因分型方法来可视化分析毒株与其他禽流感病毒之间的关系。
在分析的毒株中发现了高度的遗传多样性,因为在系统发育树中鉴定出了多个亚组。在血凝素(HA)树中,波兰毒株聚集在两个不同的亚分支中。发现聚合酶基本蛋白2(PB2)、聚合酶基本蛋白1(PB1)、聚合酶酸性蛋白(PA)和核蛋白(NP)具有高度多样性,因为可以区分出5至8个亚谱系。每个毒株都有不同的基因组合,尽管在两个分离株之间观察到多达八个基因片段中的六个存在关系。每个波兰毒株的至少一个片段与家禽分离株存在关联。
测试毒株的基因组结构表明存在广泛的重配现象,尽管可以注意到对特定基因组合的偏好。与家禽源分离株的显著关联强调了持续监测自然宿主中循环的禽流感病毒基因库的必要性。