鸽源新城疫病毒的鉴定及HN基因遗传演化分析
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摘要
根据发病鸽群的临床症状、病理剖检、组织病理学变化,并结合RT-PCR方法对武汉地区某鸽场疑似鸽新城疫进行了确诊,同时对分离的2株鸽源新城疫病毒(NDV)进行了HN基因结构特征及其与已知毒株的遗传相关性分析。结果表明:两株鸽源NDV分离株HN基因片段均含有6个保守的糖基化位点。2株NDV分离株HN基因序列与参考株同源性在80.0%~97.2%,与湖北株(AY225110.HB92)、印度株(KM056357.ndv61/B1)、江苏株(GQ245832.YC-24-07-CH)、陕西株(JF795531.HX01)在同一分支上,为基因II型。研究为武汉地区鸽新城疫的防控提供了理论依据。
Based on clinical symptoms,gross pathologic and histopathological lesions and RT-PCR,one case suspicious of Newcastle disease from pigeons in Wuhan was diagnosed. The molecular characterization and phylogenetic analysis of HN gene were performed in two strains of Newcastle disease virus( NDV). The results showed that the two virus strains contain six conservative glycosylation sites. The HN gene of two NDV isolates shared 98. 5 % to 99. 9 % of nucleotide identity with that of the reference strains. And the two isolated strains are on the same phylogenetic branch with some reference strains such as AY225110. HB92,KM056357. ndv61/B1,GQ245832. YC-24-07-CH and JF795531. HX01,which belong to genotype II. This study provides a basis for control of pigeon Newcastle disease in Wuhan.
Based on clinical symptoms,gross pathologic and histopathological lesions and RT-PCR,one case suspicious of Newcastle disease from pigeons in Wuhan was diagnosed. The molecular characterization and phylogenetic analysis of HN gene were performed in two strains of Newcastle disease virus( NDV). The results showed that the two virus strains contain six conservative glycosylation sites. The HN gene of two NDV isolates shared 98. 5 % to 99. 9 % of nucleotide identity with that of the reference strains. And the two isolated strains are on the same phylogenetic branch with some reference strains such as AY225110. HB92,KM056357. ndv61/B1,GQ245832. YC-24-07-CH and JF795531. HX01,which belong to genotype II. This study provides a basis for control of pigeon Newcastle disease in Wuhan.
引文
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[5]谷松至,王新,杜冬菊,等.鹅源新城疫病毒的分离鉴定及其致病性[J].中国兽医学报,2013,33(7):995-998.
[6]丁卫星,王荣谈,孙明昌,等.鸡新城疫疫苗预防鸽Ⅰ型副黏病毒病的免疫研究[J].上海农业学报,2014,30(1):72-74.
[7]李仕超.鸽源新城疫病毒的遗传进化分析及其免疫原性研究[D].北京:中国农业科学院,2015.
[8]朱家春.鸡新城疫及其在某鸡场鸡群发病样本的实验室检查[J].养殖技术顾问,2014(2):90-93.
[9]Ballagi P A,Wehman E,Herczeg J,et al.Identification and grouping of Newcastle disease virus strains by restriction site analysis of a region from F gene[J].Arch Virology,1996,141:243-61.
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[13]梁俊文.新城疫分离株P和HN基因分子进化与致病性的相关研究[D].济南:山东师范大学,2010.
[14]Mc Ginnes L W,Wilde A.The role of the individual cysteine resides in the formation of the mature,antigentic HN protein of Newcastle disease virus[J].Virology,1994,200:470-483.
[15]Huang Z,Panda A,Elankumaran S,et al.The hemagglutininneuraminidase protein of Newcastle disease virus determines tropism and virulence[J].Journal of Virology,2004,78(8):4176-4184.
[16]Seal B,Crawford J,Sellers H,et al.Nucleotide sequence analysis of the Newcastle disease virus nucleocapsid protein gene and phylogenetic relationships among the Paramyxoviridae[J].Virus Research,2002,83(1-2):119-129.
[2]De Leeuw O S,Koch G,Hartog L,et al.Virulence of Newcastle disease virus is determined by the cleavage site of the fusion protein and by both the stem region and globular head of the haemagglutinin-neuraminidase protein[J].Journal of General Virology,2005,86(6):1759-1769.
[3]Peeters B P,de Leeuw O S,Koch G,et al.Rescue of Newcastle disease virus from cloned c DNA:evidence that cleavability of the fusion protein is a major determinant for virulence[J].Journal of Virology,1999,73(6):5001-5009.
[4]刘栋,李建亮,秦卓明,等.NDV山东分离株的生物学特性及其HN基因的克隆与序列分析[J].微生物学通报,2006,33(4):123-128.
[5]谷松至,王新,杜冬菊,等.鹅源新城疫病毒的分离鉴定及其致病性[J].中国兽医学报,2013,33(7):995-998.
[6]丁卫星,王荣谈,孙明昌,等.鸡新城疫疫苗预防鸽Ⅰ型副黏病毒病的免疫研究[J].上海农业学报,2014,30(1):72-74.
[7]李仕超.鸽源新城疫病毒的遗传进化分析及其免疫原性研究[D].北京:中国农业科学院,2015.
[8]朱家春.鸡新城疫及其在某鸡场鸡群发病样本的实验室检查[J].养殖技术顾问,2014(2):90-93.
[9]Ballagi P A,Wehman E,Herczeg J,et al.Identification and grouping of Newcastle disease virus strains by restriction site analysis of a region from F gene[J].Arch Virology,1996,141:243-61.
[10]Chu D K,Leung C Y,Gilbert M,et al.Avian coronavirus in wild aquatic birds[J].Journal of Virology,2011,85(23):12815-12820.
[11]Kalata E F,baldauf C.Newcastle disease in freeding and pet birds.In D J Alexander(ed.).Newcastle Disease[D].Kluwer Academic Publishers:Boston,MA,197-246.
[12]Miller PJ,Kim LM,Ip HS,et al.Evolutionary dynamics of Newcastle disease virus[J].Virology,2009,391(1):64-72.
[13]梁俊文.新城疫分离株P和HN基因分子进化与致病性的相关研究[D].济南:山东师范大学,2010.
[14]Mc Ginnes L W,Wilde A.The role of the individual cysteine resides in the formation of the mature,antigentic HN protein of Newcastle disease virus[J].Virology,1994,200:470-483.
[15]Huang Z,Panda A,Elankumaran S,et al.The hemagglutininneuraminidase protein of Newcastle disease virus determines tropism and virulence[J].Journal of Virology,2004,78(8):4176-4184.
[16]Seal B,Crawford J,Sellers H,et al.Nucleotide sequence analysis of the Newcastle disease virus nucleocapsid protein gene and phylogenetic relationships among the Paramyxoviridae[J].Virus Research,2002,83(1-2):119-129.