鸽新城疫病毒BJ-C分离株基因组测序及系统发育分析

Genome sequencing and phylogenetic analysis of Newcastle disease virus BJ-C strain in pigeons

【背景】鸽新城疫是由鸽Ⅰ型副黏病毒(pigeon paramyxovirus typeⅠ,PPMV-1)感染引起的危害最严重的疫病之一,至今尚无有效的防控制剂。【目的】分析鸽新城疫病毒BJ-C株的基因组信息及系统发育关系,为鸽新城疫的防控提供科学依据。【方法】设计首尾重叠的6对特异性引物,利用分段扩增的方法,以鸽新城疫病毒BJ-C株基因组cDNA为模板,分别扩增、测序后进行全基因组序列拼接。以NCBI数据库中发布的新城疫病毒序列为参考,针对鸽新城疫病毒BJ-C株的基因组、F基因建立系统发育树。【结果】鸽新城疫病毒BJ-C株的基因组全长为15192 nt。基于全基因组的系统发育分析发现其与PPMV-1/BJ-01/CH株的系统发育关系最近,核苷酸相似性为99.96%,氨基酸相似性高达100%,属于同一个分支,而与LaSota疫苗株等其他新城疫毒株的亲缘关系相对较远。基于F基因序列的系统发育树分析发现BJ-C株F基因与我国的BJP2013株同属一个分支。ClassⅡ类Ⅵ亚型F基因高变区序列(47-420nt)比对结果显示,安徽株Pigeon/Anhui/2369/2012、广东株Pigeon/Guangdong/GZ288/2013、北京株BJP13、浙江株Pigeon/Zhejiang/2036/2012及比利时株PPMV-1/Belgium/11-09620/2011等与BJ-C毒株处于同一个分支,同属Ⅵb亚型。【结论】本研究获得了鸽新城疫病毒BJ-C株全基因组序列,分析了其系统发育关系,确定其属于ClassⅡ类Ⅵb型,为后续防控产品的开发提供了理论依据。

[Background]Pigeon Newcastle disease(ND),caused by pigeon paramyxovirus typeⅠ(PPMV-1),is a severe infectious disease.However,there is no effective method to control this disease.[Objective]To elucidate the phylogenetic analysis of BJ-C and provide a scientific basis for the prevention and control of pigeon ND.[Methods]Six pairs of specific primers overlapped at the beginning and ending were designed.The genomic cDNA sequence of BJ-C strain was used as the template for amplification and sequencing.The phylogenetic tree of BJ-C strain was established by sequence alignment with the published PPMV-1 and NDV strains on the NCBI website.[Results]The genome length of BJ-C strain was 15192 nt.BJ-C strain had the closest genetic relationship with PPMV-1/BJ-01/CH strain based on the whole genome analysis,with nucleotide similarity of 99.96%and amino acid similarity of 100%.BJ-C strain shared the same branch with PPMV-1/BJ-01/CH strain,whereas it was far from other ND virus strains such as the LaSota vaccine strain.The phylogenetic tree built based on F gene showed that BJ-C strain was in the same branch with BJP2013 strain isolated in China.The sequence alignment of the hypervariable region(47–420 nt)of F gene showed that theⅥb subtype strains including Pigeon/Anhui/2369/2012,Pigeon/Guangdong/GZ288/2013,BJP13,Pigeon/Zhejiang/2036/2012,PPMV-1/Belgium/11-09620/2011,and BJ-C were in the same clade.[Conclusion]The full-length genome sequence of BJ-C strain was obtained and the strain was grouped to the classⅡⅥb type based on phylogenetic analysis.Our results are of theoretical significance for the development of pigeon ND vaccines and other prevention and control methods.