鹅细小病毒DY16株的分子鉴定及重组分析
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摘要
为了了解鹅细小病毒(goose parvovirus,GPV)田间分离株是否存在变异可能,本研究对GPV分离株DY16进行了全基因组测序和重组分析。DY16株基因组由5 046个碱基组成,与国内外的5个强毒株同源性在98.1%以上。末端倒置重复(inverted terminal repeats,ITR)由414个碱基组成,与已报道的多个GPV强毒株同样在ITR的回文区茎部存在14个碱基对缺失。Simplot软件在DY16株VP1基因内部检测到2个重组信号,序列比对显示DY16株位于重组第1区域的10个和重组第2区域8个核苷酸位点分别与匈牙利的B株和国内弱毒疫苗株SYG61v相一致,而不同于先前的国内强毒株,但均未产生氨基酸改变。不同来源毒株在VP1基因内部重组对于GPV致病性的影响有待深入探究。
In order to dissect whether there are goose parvovirus(GPV) variants prevailing in the field,in this study,the genome of a GPV isolate DY16 was sequenced and recombination analysis was executed.The DY16 genome is comprised of 5 046 nucleotides,which displays over 98.1% nucleotide homology with five virulent strains from China and Hungary.The inverted terminal repeats(ITR) consists of 414 nucleotides,and harbors a 14-nucleotide-pair deletions in the stem of the palindromic region as observed in other GPV strains.Two recombination signals were detected in the VP1 gene of DY16 by using the Simplot software.Ten nucleotides in the first recombination site of DY16 and eight nucleotides in the second recombination site were in accordance with those of strain B from Hungary and the attenuated vaccine strain SYG61 v respectively,but different from those of the virulent strains previously characterized from China.However,all the nucleotide changes occurred in the recombination region of strain DY16 do not result in amino acid alterations.Whether the recombination occurred in the VP1 gene between the GPV strains with various geographic origins and virulence can contribute to the pathogenic change deserves to be thoroughly investigated in the future.
In order to dissect whether there are goose parvovirus(GPV) variants prevailing in the field,in this study,the genome of a GPV isolate DY16 was sequenced and recombination analysis was executed.The DY16 genome is comprised of 5 046 nucleotides,which displays over 98.1% nucleotide homology with five virulent strains from China and Hungary.The inverted terminal repeats(ITR) consists of 414 nucleotides,and harbors a 14-nucleotide-pair deletions in the stem of the palindromic region as observed in other GPV strains.Two recombination signals were detected in the VP1 gene of DY16 by using the Simplot software.Ten nucleotides in the first recombination site of DY16 and eight nucleotides in the second recombination site were in accordance with those of strain B from Hungary and the attenuated vaccine strain SYG61 v respectively,but different from those of the virulent strains previously characterized from China.However,all the nucleotide changes occurred in the recombination region of strain DY16 do not result in amino acid alterations.Whether the recombination occurred in the VP1 gene between the GPV strains with various geographic origins and virulence can contribute to the pathogenic change deserves to be thoroughly investigated in the future.
引文
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[2] 王艳秋,张紫璇,高旭,等.鹅细小病毒TaqMan荧光定量PCR方法的建立及临床应用[J].中国兽医学报,2018,38(6):1100-1104.
[3] 方定一.小鹅瘟的介绍[J].中国畜牧兽医,1962(8):19-20.
[4] 方定一,王永坤,郑玉美,等.小鹅瘟病原体及其特异防治的研究[J].中国农业科学,1981(4):1-7.
[5] DERZSY D.A viral disease of goslings.I.epidemiological,clinical,pathological and etiological studies[J].Acta Zool Acad Sci H,1967,17:443-448.
[6] 周阳生,田慧芳,过建寅,等.小鹅瘟疫苗对初生雏鹅的安全性及免疫力试验[J].中国兽医杂志,1984(9):2-4.
[7] 王永坤.小鹅瘟的诊断与防制[M].1版.南京:江苏科学技术出版社,1992(6):29-38.
[8] WANG J,DUAN J,ZHU L,et al.Sequencing and generation of an infectious clone of the pathogenic goose parvovirus strain LH[J].Arch Virol,2005,160(3):711-718.
[9] SHIEN J,WANG Y,CHEN C,et al.Identification of sequence changes in live attenuated goose parvovirus vaccine strains developed in Asia and Europe[J].Avian Pathol,2008,37(5):499-505.
[10] 郑玉美,李俊宝,周阳生.中国和匈牙利小鹅瘟病毒的交叉中和试验[J].江苏农学院学报,1985,6(4):45-48.
[11] WANG J,DUAN J,MENG X,et al.Cloning of the genome of a goose parvovirus vaccine strain SYG61v and rescue of infectious virions from recombinant plasmids in embryonated goose eggs[J].J Virol Methods,2014,200:41-46.
[12] 王建业,黄钰,段进坤,等.鹅细小病毒鹅胚化疫苗株SYG61v对雏鹅毒力减弱的遗传基础分析[J].中国动物传染病学报,2015,23(4):19-24.
[13] WANG J,LING J,WANG Z,et al.Molecular characterization of a novel Muscovy duck parvovirus isolate:evidence of recombination between classical MDPV and goose parvovirus strains[J].BMC Vet Res,2017,13(1):327.