一株新亚群禽白血病病毒全基因组序列分析

摘要

为了解外源性禽白血病病毒(ALV)在山东省部分地区肉鸡中的流行状况,采集无菌抗凝血分离血浆接种DF-1细胞、ELISA p27抗原检测以及DNA提取进行PCR扩增等方法,从山东省某地区大型养殖场不同个体养殖场出栏肉鸡群中分离鉴定出1株ALV,命名为FC1505.分析其囊膜糖蛋白gp85氨基酸序列与近年来从地方品种鸡中疑似新亚群ALV-K分离株的相似性最高,相似性均在95％以上,而与其他已知亚群ALV的相似性均低于90％.为了进一步分析该分离株分子特性,对其进行全基因组测序,并与已知亚群ALV分离株序列进行比较.结果表明,FC1505分离株整个基因组中gag、pol和gp37基因相对保守,与ALV参考毒株序列相似性都在90％以上,但均与疑似K业群的ALV分离株相似性最高;全基因组序列分析进一步说明FC1505属于ALV-K.本研究继我国江苏省和华南地区K亚群ALV报道后,首次从山东地区肉鸡中鉴定到一株ALV-K并完成其全基因组序列分析.%In order to understand the epidemic situation of exogenous avian leukemia virus (ALV)in broiler flocks in Shandong province,blood plasma were collected for virus isolation and identification by ELISA (detection of P27 antigen) and PCR.A strain of new ALV subgroup was isolated and identified from commercial broiler flocks in a farm of Shandong province,named FC1505.The env gene of the isolate was sequenced and indicated that the amino acid sequence of gp85 had the highest gp85 identity of more than 95％ to the strains of suspected subgroup K of ALV from Chinese native breed chickens.Its gp85 identity to other known chicken ALV subgroups A-E and J was lower than 90 ％.In order to further analyze the molecular properties of the isolate,the genome of the virus was sequenced and compared with those from other ALV reference strain of different subgroups.The results showed that the gag,pol and gp37 genes of the isolate were relatively conservative,with the identity more than 90％,and showed the highest identity to suspected ALV-K isolates.Whole genome sequence analysis supported that the FC1505 isolate belongs to ALV-K.Following the reports of K subgroup ALV in Jiangsu and Southern China provinces of China,this is the first report of an ALV-K isolate from broilers of Shandong province and its whole genome analysis.