黄羽父母代种鸡混合感染禽白血病病毒与马立克病病毒的鉴定
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摘要
广西某养殖场130日龄父母代种鸡发生临床肿瘤样病变和死亡,对病鸡采用病理解剖、组织病理学观察、PCR检测、病毒分离以及分离株重要基因的序列测定和病原鉴定。结果显示:病鸡的心脏、肝脏、脾脏等部位表现有肿瘤样病变; PCR检测及病毒分离培养有J亚群禽白血病病毒(avian leukosis virus subgroup J,ALV-J)和马立克病病毒(Marek's disease virus,MDV)的感染; ALV-J分离株env基因的序列与10株ALV-J参考株的核苷酸相似性为87. 2%～97. 7%,与ALV A-E亚型参考株的相似性为53. 5%～54. 7%;与ALV-J英国原型株HPRS-103的env基因糖基化位点进行分析比较,部分糖基化位点发生了改变; MDV分离株meq基因序列与8株MDV参考株的核苷酸相似性为98. 7%～99. 4%,分离株在第71～80位氨基酸发生突变,符合国内强毒分离株的特征。结果说明:该鸡群为ALV-J和MDV的混合感染。
A parent stock of breeding chickens on a farm in Guangxipresented clinical tumor-like lesions and suffered from death at 130 days of age. Pathological anatomy,histopathological observation,PCR detection,virus isolation,and sequencing and analysis of important genes of isolates from the virus of the diseased chickens were performed for diagnosis. The results showed that the heart,liver,spleen and other organs of the diseased chickens showed tumor-like gross lesions. PCR detection and virus isolation of the cell cultures showed co-infection of avian leukosis virus subgroup J( ALV-J) and Marek's disease virus( MDV). The nucleotide sequence of the env gene of the ALV-J isolate was 87. 2%-97. 7% similar to those of the 10 ALV-J reference strains and 53. 5%-54. 7% similar to those of the ALV A-E subtype reference strains. Comparison of the glycosylation sites with the British ALV-J prototype strain HPRS-103 revealed changes of some glycosylation sites. The meq gene nucleotide sequence of the MDV isolate was 98. 7%-99. 4% similar with 8 MDV reference strains and the mutations at the region of No. 71-80 amino acids were found,which was consistent with the characteristics of the domestic virulent MDV isolates( v MDV). The results obtained here indicated that the flock of the studied chickens was co-infected with ALV-J and MDV.
A parent stock of breeding chickens on a farm in Guangxipresented clinical tumor-like lesions and suffered from death at 130 days of age. Pathological anatomy,histopathological observation,PCR detection,virus isolation,and sequencing and analysis of important genes of isolates from the virus of the diseased chickens were performed for diagnosis. The results showed that the heart,liver,spleen and other organs of the diseased chickens showed tumor-like gross lesions. PCR detection and virus isolation of the cell cultures showed co-infection of avian leukosis virus subgroup J( ALV-J) and Marek's disease virus( MDV). The nucleotide sequence of the env gene of the ALV-J isolate was 87. 2%-97. 7% similar to those of the 10 ALV-J reference strains and 53. 5%-54. 7% similar to those of the ALV A-E subtype reference strains. Comparison of the glycosylation sites with the British ALV-J prototype strain HPRS-103 revealed changes of some glycosylation sites. The meq gene nucleotide sequence of the MDV isolate was 98. 7%-99. 4% similar with 8 MDV reference strains and the mutations at the region of No. 71-80 amino acids were found,which was consistent with the characteristics of the domestic virulent MDV isolates( v MDV). The results obtained here indicated that the flock of the studied chickens was co-infected with ALV-J and MDV.
引文
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[14]王培坤,毕玉彧,秦丽莉,等.三黄鸡临床血管瘤病例中分离出A亚群与J亚群禽白血病病毒[J].动物医学进展,2015,36(3):13-16.
[15]李海娟,林璐璐,王培坤,等.一株引起90日龄三黄鸡髓细胞瘤病ALV-J的分离与鉴定[J].中国家禽,2017,39(14):61-64.
[16]施维松,刘长军,张艳萍,等. 4株鸡马立克氏病病毒国内分离株meq基因的克隆与序列分析[J].病毒学报,2008,24(2):117-125.
[17] Tian M,Zhao Y,Lin Y,et al. Comparative analysis of oncogenic genes revealed unique evolutionary features of field Marek's disease virus prevalent in recent years in China[J]. Virol J,2011,8(1):1-11.
[18]韦平,崔治中,Lee L F.马立克氏病病毒meq基因功能研究[J].广西科学,2003,10(1):52-62.
[19] Shamblin C E,Greene N,Arumugaswami V,et al. Comparative analysis of Marek's disease virus(MDV)glycoprotein-,lytic antigen pp38-and transformation antigen Meq-encoding genes:association of meq mutations with MDVs of high virulence[J]. Vet Microbiol,2004,102(3):147-167.
[20]成子强,张玲娟,刘杰,等.蛋鸡中发现J亚群白血病与网状内皮增生症自然混合感染[J].中国兽医学报,2006,26(6):586-590.
[2]韦平,丁家波.几种引起家禽免疫抑制的病毒性疾病及其作用机理[J].中国预防兽医学报,2000,22(4):316-318.
[3] 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.
[4]蔡宝祥.家畜传染病学[M].北京:北京中国农业出版社,2001.
[5]周庚寅.组织病理学技术[M].北京:北京大学医学出版社,2006.
[6]韦平,崔治中.马立克氏病病毒不同致病型meq基因的比较研究[J].中国预防兽医学报,2002,24(2):88-92.
[7] Silva R F. Differentiation of pathogenic and non-pathogenic serotype1 Marek's disease viruses(MDVs)by the polymerase chain reaction amplification of the tandem direct repeats within the MDV genome[J]. Avian Dis,1992,36(3):521.
[8]朱玉贤,李毅,郑晓峰.现代分子生物学[M].北京:高等教育出版社,2007.
[9] Payne L N,Brown S R,Bumstead N,et al. A novel subgroup of exogenous avian leukosis virus in chickens[J]. J Gen Virol,1991,72(Pt 4)(4):801.
[10]张志,崔治中,赵宏坤,等.商品代肉鸡J亚群禽白血病的病理及病毒分离鉴定[J].中国兽医杂志,2002,38(6):6-8.
[11] Rispens B H,Van V H,Mastenbroek N,et al. Control of Marek's disease in the Netherlands. I. Isolation of an avirulent Marek's disease virus(strain CVI988)and its use in laboratory vaccination trials[J]. Avian Dis,1972,16(1):108-125.
[12]滕丽琼,韦平,宋中宝,等.一株整合有禽反转录病毒长末端重复序列的马立克氏病病毒野毒株的致病性研究[J].病毒学报,2009(5):376-381.
[13]申艳玮.选择压作用下J亚群禽白血病病毒突变位点的研究[D].泰安:山东农业大学,2014.
[14]王培坤,毕玉彧,秦丽莉,等.三黄鸡临床血管瘤病例中分离出A亚群与J亚群禽白血病病毒[J].动物医学进展,2015,36(3):13-16.
[15]李海娟,林璐璐,王培坤,等.一株引起90日龄三黄鸡髓细胞瘤病ALV-J的分离与鉴定[J].中国家禽,2017,39(14):61-64.
[16]施维松,刘长军,张艳萍,等. 4株鸡马立克氏病病毒国内分离株meq基因的克隆与序列分析[J].病毒学报,2008,24(2):117-125.
[17] Tian M,Zhao Y,Lin Y,et al. Comparative analysis of oncogenic genes revealed unique evolutionary features of field Marek's disease virus prevalent in recent years in China[J]. Virol J,2011,8(1):1-11.
[18]韦平,崔治中,Lee L F.马立克氏病病毒meq基因功能研究[J].广西科学,2003,10(1):52-62.
[19] Shamblin C E,Greene N,Arumugaswami V,et al. Comparative analysis of Marek's disease virus(MDV)glycoprotein-,lytic antigen pp38-and transformation antigen Meq-encoding genes:association of meq mutations with MDVs of high virulence[J]. Vet Microbiol,2004,102(3):147-167.
[20]成子强,张玲娟,刘杰,等.蛋鸡中发现J亚群白血病与网状内皮增生症自然混合感染[J].中国兽医学报,2006,26(6):586-590.
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