一株鸡传染性贫血病毒的分离鉴定及全基因组序列分析
|
摘要
为分离鉴定一株鸡传染性贫血病毒(CAV),以及了解其基因组特征,本实验利用MDCC-MSB1细胞对黑龙江某鸡场疑似患病鸡的肝脏组织进行病毒分离,经PCR和间接免疫荧光试验鉴定,结果显示分离得到一株CAV,命名为HLJ16069。克隆该病毒株全基因组,测序拼接后获得全长序列,将该病毒株序列与国内外已发表的其它病毒全基因组序列进行同源比对和进化分析,结果显示同源性达96.0%以上,亲缘关系最远的是中国分离株SD1403,而与日本分离株C369亲缘关系最近(99.1%);氨基酸序列分析显示,HLJ16069在VP1的75、89、141和394位氨基酸存在有意义的突变。本研究发现了一些与CAV毒力相关的分子特征,为CAV的毒力变化提供了数据支持。
In order to investigate the characteristic of whole genome of prevalent chicken anemia virus(CAV), a CAV was isolated from the liver sample of suspected illness breeder chicken which was collected in Heilongjiang province by passaging in MDCC-MSB1 cells. The virus was identified as CAV based on specific PCR and indirect immuno-fluorescence, named HLJ16069.The whole genome of the isolate was amplified and then was assembled for sequence alignment with other sequences in Gen Bank.The whole genome sequence of HLJ16069 shared 96.0%-99.1% identity of nucleotide sequences with those CAV reference strains, the minimum difference of strain is C369 of Japan. Amino acids alignment showed a series of virulent related changes at aa75, aa89, aa141 and aa394 of VP1. This study alerts the serious economic threat to the poultry industry, and also revealed some molecular characteristics on virulence of CAV.
In order to investigate the characteristic of whole genome of prevalent chicken anemia virus(CAV), a CAV was isolated from the liver sample of suspected illness breeder chicken which was collected in Heilongjiang province by passaging in MDCC-MSB1 cells. The virus was identified as CAV based on specific PCR and indirect immuno-fluorescence, named HLJ16069.The whole genome of the isolate was amplified and then was assembled for sequence alignment with other sequences in Gen Bank.The whole genome sequence of HLJ16069 shared 96.0%-99.1% identity of nucleotide sequences with those CAV reference strains, the minimum difference of strain is C369 of Japan. Amino acids alignment showed a series of virulent related changes at aa75, aa89, aa141 and aa394 of VP1. This study alerts the serious economic threat to the poultry industry, and also revealed some molecular characteristics on virulence of CAV.
引文
[1]Yuasa N,Yoshida I.Isolation and Some Characteristics of an Agent Inducing Anemia in Chicks[J].Avian Dis,1979,23(2):366-385.
[2]Varela A P,Dos Santos H F,Cibulski S P,et al.Chicken anemia virus and avian gyrovirus 2 as contaminants in poultry vaccines[J].Biologicals,2014,42(6):346-350.
[3]Ducatez M F,Chen H,Guan Y,et al.Molecular epidemiology of chicken anemia virus(CAV)in Southeastern Chinese live birds markets[J].Avian Dis,2008,52:68-73.
[4]Olszewska-Tomczyk M,SwietońE,Minta Z,et al.Occurrence and Phylogenetic Studies of Chicken Anemia Virus from Polish Broiler Flocks[J].Avian Dis,2016,60(1):70-74.
[5]Kucharski T J,Ng T F,Sharon D M,et al.Activation of the Chicken Anemia Virus Apoptin Protein by Chk1/2 Phosphorylation Is Required for Apoptotic Activity and Efficient Viral Replication[J].Virol,2016,90(20):9433-9445.
[6]Kucharski T J,Ng T F,Sharon D M,et al.Activation of the Chicken Anemia Virus Apoptin Protein by Chk1/2 Phosphorylation Is Required for Apoptotic Activity and Efficient Viral Replication[J].J Virol,2016,90(20):9433-9445.
[7]Todd D,Connor T J,Creelan J L,et al.Effect of multiple cell culture passages on the biological behaviour of chicken anaemia virus[J].Avian Pathol,1998,27(1):74-79.
[8]Todd D,Scott A N,Ball N W,et al.Molecular basis of the attenuation exhibited by molecularly cloned highly passaged chicken anemia virus isolates[J].J Virol,2002,76(76):8472-8474.
[9]Chowdhury S M Z H,Omar A R,Aini I,et al.Pathogenicity,sequence and phylogenetic analysis of Malaysian Chicken anaemia virus,obtained after low and high passages in MSB-1cells[J].Arch Virol,2003,148(148):2437-2448.
[2]Varela A P,Dos Santos H F,Cibulski S P,et al.Chicken anemia virus and avian gyrovirus 2 as contaminants in poultry vaccines[J].Biologicals,2014,42(6):346-350.
[3]Ducatez M F,Chen H,Guan Y,et al.Molecular epidemiology of chicken anemia virus(CAV)in Southeastern Chinese live birds markets[J].Avian Dis,2008,52:68-73.
[4]Olszewska-Tomczyk M,SwietońE,Minta Z,et al.Occurrence and Phylogenetic Studies of Chicken Anemia Virus from Polish Broiler Flocks[J].Avian Dis,2016,60(1):70-74.
[5]Kucharski T J,Ng T F,Sharon D M,et al.Activation of the Chicken Anemia Virus Apoptin Protein by Chk1/2 Phosphorylation Is Required for Apoptotic Activity and Efficient Viral Replication[J].Virol,2016,90(20):9433-9445.
[6]Kucharski T J,Ng T F,Sharon D M,et al.Activation of the Chicken Anemia Virus Apoptin Protein by Chk1/2 Phosphorylation Is Required for Apoptotic Activity and Efficient Viral Replication[J].J Virol,2016,90(20):9433-9445.
[7]Todd D,Connor T J,Creelan J L,et al.Effect of multiple cell culture passages on the biological behaviour of chicken anaemia virus[J].Avian Pathol,1998,27(1):74-79.
[8]Todd D,Scott A N,Ball N W,et al.Molecular basis of the attenuation exhibited by molecularly cloned highly passaged chicken anemia virus isolates[J].J Virol,2002,76(76):8472-8474.
[9]Chowdhury S M Z H,Omar A R,Aini I,et al.Pathogenicity,sequence and phylogenetic analysis of Malaysian Chicken anaemia virus,obtained after low and high passages in MSB-1cells[J].Arch Virol,2003,148(148):2437-2448.