J亚群禽白血病病毒GD14A8株分离鉴定及遗传进化分析
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摘要
为了解J亚群禽白血病病毒(ALV-J)的流行和变异情况,本研究利用ALV-p27群特异性抗原检测、PCR鉴定、病毒分离和全基因组测序分析等方法,从广东某鸡场分离鉴定出1株ALV-J,命名为GD14A8。全基因组序列分析表明,GD14A8分离株的gag、pol、env基因和LTR相对保守,而3′UTR变异较大,与ALV-J参考毒株序列同源性仅为82.5%~92.5%,其中rTM和E元件呈现大量碱基缺失,U3区出现与血管瘤分离株NHH和SCDY1类似的11个碱基缺失,可能与血管瘤形成相关。遗传进化分析表明,GD14A8分离株与我国鸡源ALV-J血管瘤分离株SCDY1亲缘关系最近。本研究为分析黄羽肉鸡源ALV-J毒株的分子特征提供了重要的数据资料,为深入了解广东地区ALV-J的变异趋势提供参考。
To investigate the prevalence of subgroup J avian leukosis virus(ALV-J)in yellow broilers,one subgroup ALV-J named GD14A8 was isolated from yellow broiler in a farm of Guangdong province using ELISA(p27antigen detection),PCR and genomic sequencing assays.Genome analysis showed that gag,pol,envgenes and LTR region of the isolate were highly conserved,while 3′UTR displayed many variation,and the rTM region and E element developed many nucleotide deletion.Like other ALV-J NHH and SCDY1 strain,11bp deletion of U3 region appeared in the genome of GD14A8 isolate,which might be associated with hemangioma formation.Phylogenetic analysis showed that the isolate GD14A8 strain was closely identical to the SCDY1 strain.All these data provide a scientific basis of the molecular characteristic of ALV-J.
To investigate the prevalence of subgroup J avian leukosis virus(ALV-J)in yellow broilers,one subgroup ALV-J named GD14A8 was isolated from yellow broiler in a farm of Guangdong province using ELISA(p27antigen detection),PCR and genomic sequencing assays.Genome analysis showed that gag,pol,envgenes and LTR region of the isolate were highly conserved,while 3′UTR displayed many variation,and the rTM region and E element developed many nucleotide deletion.Like other ALV-J NHH and SCDY1 strain,11bp deletion of U3 region appeared in the genome of GD14A8 isolate,which might be associated with hemangioma formation.Phylogenetic analysis showed that the isolate GD14A8 strain was closely identical to the SCDY1 strain.All these data provide a scientific basis of the molecular characteristic of ALV-J.
引文
[1]秦建如,冯敏,郝建勇,等.3个江西地方鸡品种的禽白血病病毒病原学调查[J].中国兽医学报,2015,35(9):1452-1455.
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[10]QU Y,SUN H,SUN M,et al.Sequence analysis for the complete proviral genome of avian leukosis virus subgroup J associated with haemangiomas,leiomyosarcomas and myelomas in layer flocks[J].Acta Vet Hung,2012,60(3):343-354.
[11]HAN C,HAO R,LIU L,et al.Molecular characterization of 3'UTRs of J subgroup avian leukosis virus in passerine birds in China[J].Arch Virol,2015,160(3):845-849.
[12]CHESTERS P M,SMITH L P,NAIR V.E(XSR)element contributes to the oncogenicity of avian leukosis virus(subgroup J)[J].J Gen Virol,2006,87:2685-2692.
[13]GAO Y,YUN B,QIN L,et al.Molecular epidemiology of avian leukosis virus subgroup J in layer flocks in China[J].J Clin Microbiol,2012,50(3):953-960.
[14]SHI M,TIAN M,LIU C,et al.Sequence analysis for the complete proviral genome of subgroup J avian leukosis virus associated with hemangioma:a special 11bp deletion was observed in U3region of 3′UTR[J].Virol J,2011(8):158.
[15]JI X,WANG Q,LI X,et al.A 19-nucleotide insertion in the leader sequence of avian leukosis virus subgroup J contributes to its replication in vitro but is not related to its pathogenicity in vivo[J].PLoS One,2014,9(1):84797.
[16]吴轶娜,陆建平,郑肖利.磁性纳米靶向Apo-AⅠ基因治疗对大鼠动脉粥样硬化斑块发展的作用[J].生物医学工程与临床,2009,13(3):235-240.
[2]PAYNE L N,BROWN S R,BUMSTE A N,et al.A novel subgroup of exogenous avian leukosis virus in chickens[J].J Gen Virol,1991,72(Pt 4):801-807.
[3]杜岩,崔治中,秦爱建,等.鸡的J亚群白血病病毒的分离及部分序列比较[J].病毒学报,2000,16(4):341-346.
[4]李艳,崔治中,孙淑红.黄羽肉鸡J亚群白血病病毒的分子生物学特性和致病性[J].病毒学报,2007,23(3):207-211.
[5]潘孝成,赵瑞宏,胡晓苗,等.黄羽鸡J亚群禽白血病病毒的分离及gp85基因分析[J].动物医学进展,2014,34(2):51-54.
[6]张丹俊,赵瑞宏,沈学怀,等.黄羽肉鸡J亚群禽白血病肿瘤及相关病变发生规律的研究[J].中国家禽,2013,35(20):15-19.
[7]SMITH L M,BROWN S R,HOWES K,et al.Development and application of polymerase chain reaction(PCR)tests for the detection of subgroup J avian leukosis virus[J].Virus Res,1998,54(1):87-98.
[8]BIETH E,DARLIX J L.Complete nucleotide sequence of a highly infectious avian leukosis virus[J].Nucleic Acids Res,1992,20(2):367.
[9]SILVA R F,FADLY A M,TAYLOR S P.Development of a polymerase chain reaction to differentiate avian leukosis virus(ALV)subgroups:detection of an ALV contaminant in commercial Marek's disease vaccines[J].Avian Dis,2007,51(3):663-667.
[10]QU Y,SUN H,SUN M,et al.Sequence analysis for the complete proviral genome of avian leukosis virus subgroup J associated with haemangiomas,leiomyosarcomas and myelomas in layer flocks[J].Acta Vet Hung,2012,60(3):343-354.
[11]HAN C,HAO R,LIU L,et al.Molecular characterization of 3'UTRs of J subgroup avian leukosis virus in passerine birds in China[J].Arch Virol,2015,160(3):845-849.
[12]CHESTERS P M,SMITH L P,NAIR V.E(XSR)element contributes to the oncogenicity of avian leukosis virus(subgroup J)[J].J Gen Virol,2006,87:2685-2692.
[13]GAO Y,YUN B,QIN L,et al.Molecular epidemiology of avian leukosis virus subgroup J in layer flocks in China[J].J Clin Microbiol,2012,50(3):953-960.
[14]SHI M,TIAN M,LIU C,et al.Sequence analysis for the complete proviral genome of subgroup J avian leukosis virus associated with hemangioma:a special 11bp deletion was observed in U3region of 3′UTR[J].Virol J,2011(8):158.
[15]JI X,WANG Q,LI X,et al.A 19-nucleotide insertion in the leader sequence of avian leukosis virus subgroup J contributes to its replication in vitro but is not related to its pathogenicity in vivo[J].PLoS One,2014,9(1):84797.
[16]吴轶娜,陆建平,郑肖利.磁性纳米靶向Apo-AⅠ基因治疗对大鼠动脉粥样硬化斑块发展的作用[J].生物医学工程与临床,2009,13(3):235-240.
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