一株鸽Ⅰ型副粘病毒的分离鉴定及F基因遗传进化分析
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摘要
东北某动物园鸽群发生异常死亡,采集病死鸽脏器,通过SPF鸡胚进行病毒分离,采用血凝试验和RT-PCR试验,鉴定结果显示尿囊液呈Ⅰ型副粘病毒核酸阳性。通过对F基因进行测序和遗传进化分析,结果表明,分离株BS0904裂解位点基序为112RRQKRF117,符合NDV强毒株的典型分子特征;分离株F基因与疫苗株LaS ota、B1、Clone 30、VG/GA、V-4、Mukteswar及经典强毒株F48E9的核苷酸同源性较低,仅为84. 2%—85. 8%;而与2011—2013年在中国多个地区鸽中所分离毒株核苷酸同源性则高达98. 8%—99. 9%,并且在遗传进化树上位于同一分支,属于ClassⅡ,基因型为Ⅵb-EU/re。该基因型毒株在中国鸽群中有较长时间、较大范围的流行传播,且与疫苗株遗传进化关系较远,说明疫苗株不能对鸽群提供有效的免疫保护,因此为了更好地对鸽群进行Ⅰ型副粘病毒的有效防控,需要针对Ⅵb基因型研发新疫苗。
Abnormal death of pigeons was documented in a zoo in northeast China,a pigeon paramyxovirus serotypeⅠ( PPMV-1) isolate was recovered from samples collected from dead pigeons and initial isolation of the virus was performed in specific-pathogen-free( SPF) chicken embryos.The isolate was identified by the hemagglutination( HA) test and RT-PCR. Sequencing and phylogenetic analysis of F gene indicated that the cleavage sites of the BS0904 isolate was112 RRQKRF117 motif in accordance with characteristic of velogenic strains. The isolate shared84. 2%-85. 8% nucleotide identity with the vaccine strains LaS ota,B1,Clone 30,VG/GA,V-4,Mukteswar,and high virulent strain F48 E9. But the isolate showed a high level of nucleotide identity( 98. 8%-99. 9%) with isolates obtained from pigeons in regions of China from 2011 to2013. It also aligned into the same sub-branch in the phylogenetic tree,which belongs to ClassⅡ,genotype Ⅵb-EU/re. The genotype strains had spread among pigeons in China for a long time and over a wide geographic range. They exhibited long distance from the vaccine stains,therefore,new vaccines phylogenetically close to the prevalent genotype Ⅵb PPMV-1 should be developed to prevent the PPMV-1 infection.
Abnormal death of pigeons was documented in a zoo in northeast China,a pigeon paramyxovirus serotypeⅠ( PPMV-1) isolate was recovered from samples collected from dead pigeons and initial isolation of the virus was performed in specific-pathogen-free( SPF) chicken embryos.The isolate was identified by the hemagglutination( HA) test and RT-PCR. Sequencing and phylogenetic analysis of F gene indicated that the cleavage sites of the BS0904 isolate was112 RRQKRF117 motif in accordance with characteristic of velogenic strains. The isolate shared84. 2%-85. 8% nucleotide identity with the vaccine strains LaS ota,B1,Clone 30,VG/GA,V-4,Mukteswar,and high virulent strain F48 E9. But the isolate showed a high level of nucleotide identity( 98. 8%-99. 9%) with isolates obtained from pigeons in regions of China from 2011 to2013. It also aligned into the same sub-branch in the phylogenetic tree,which belongs to ClassⅡ,genotype Ⅵb-EU/re. The genotype strains had spread among pigeons in China for a long time and over a wide geographic range. They exhibited long distance from the vaccine stains,therefore,new vaccines phylogenetically close to the prevalent genotype Ⅵb PPMV-1 should be developed to prevent the PPMV-1 infection.
引文
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[7]田志革.野生水禽副粘病毒的分离鉴定及Ⅴ蛋白对抗宿主细胞先天免疫机制的研究[D].哈尔滨:东北林业大学,2015.
[8]熊晓妍,孙兴臣,李敏婕,等.赛鸽新城疫病毒南京分离株的生物学特性及其F和HN基因的分析[J].中国兽医科学,2017,47(11):1363-1370.
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[10]吴双,王伟伟,曹军平,等.不同基因型新城疫病毒对美国白羽王鸽的致病性研究[J].中国预防兽医学报,2010,32(6):424-427.
[11]杨吉才,石国芳.鸽新城疫最新研究进展[J].中国动物保健,2005(9):34-36.
[12]UJVARI D,WEHMANN E,KALETA E F,et al.Phylogenetic analysis reveals extensive evolution of avian paramyxovirus type 1strains of pigeons(Columba livia)and suggests multiple species transmission[J].Virus Research,2003,96(1/2):63-73.
[13]赵振振,朱杰,许海旭,等.2011-2013年7株鸽源新城疫病毒的生物学特性鉴定及遗传进化分析[J].中国兽医学报,2016,36(11):1853-1857.
[2]ALEXANDER D J.Avian Paramyxoviridae-recent developments[J].Veterinary Microbiology,1990,23(1/4):103-114.
[3]ALEXANDER D J,RUSSELL P H,PARSONS G,et al.Antigenic and biological characterisation of avian paramyxovirus type I isolates from pigeons-an international collaborative study[J].Avian Pathology,1985,14(3):365-376.
[4]KALETA E F,ALEXANDER D J,RUSSELL P H.The first isolation of the avian PMV-1 virus responsible for the current panzootic in pigeons[J].Avian Pathology,1985,14(4):553-557.
[5]BIANCIFIORI F,FIORONI A.An occurrence of Newcastle disease in pigeons:virological and serological studies on the isolates[J].Comparative Immunology,Microbiology and Infectious Diseases,1983,6(3):247-252.
[6]LOMNICZI B,WEHMANN E,HERCZEG J,et al.Newcastle disease outbreaks in recent years in western Europe were caused by an old(VI)and a novel genotype(VII)[J].Archives of Virology,1998,143(1):49-64.
[7]田志革.野生水禽副粘病毒的分离鉴定及Ⅴ蛋白对抗宿主细胞先天免疫机制的研究[D].哈尔滨:东北林业大学,2015.
[8]熊晓妍,孙兴臣,李敏婕,等.赛鸽新城疫病毒南京分离株的生物学特性及其F和HN基因的分析[J].中国兽医科学,2017,47(11):1363-1370.
[9]李德厚,刘文灿,陈琨,等.从进口种鸽中多次检出鸽新城疫病[J].中国兽医杂志,1988,14(3):54-55.
[10]吴双,王伟伟,曹军平,等.不同基因型新城疫病毒对美国白羽王鸽的致病性研究[J].中国预防兽医学报,2010,32(6):424-427.
[11]杨吉才,石国芳.鸽新城疫最新研究进展[J].中国动物保健,2005(9):34-36.
[12]UJVARI D,WEHMANN E,KALETA E F,et al.Phylogenetic analysis reveals extensive evolution of avian paramyxovirus type 1strains of pigeons(Columba livia)and suggests multiple species transmission[J].Virus Research,2003,96(1/2):63-73.
[13]赵振振,朱杰,许海旭,等.2011-2013年7株鸽源新城疫病毒的生物学特性鉴定及遗传进化分析[J].中国兽医学报,2016,36(11):1853-1857.