贵州省鸭源新城疫病毒强毒株的遗传变异分析及致病性研究

doi:doi: 10.11843/j.issn.0366-6964.2016.08.013

畜牧兽医学报

贵州省鸭源新城疫病毒强毒株的遗传变异分析及致病性研究

段志强^1，2，许厚强^1，2，嵇辛勤^1，2，陈强¹，胡炎¹，胡顺林³，刘秀梵^3*

（1.贵州大学动物科学学院，贵阳 550025；2.贵州大学高原山地动物遗传育种与繁殖省部共建教育部重点实验室，贵阳 550025；3.扬州大学农业部畜禽传染病学重点开放实验室，扬州 225009）

收稿日期:2016-03-23 出版日期:2016-08-23 发布日期:2016-09-08
通讯作者: 刘秀梵，Tel: 0514-87991416；E-mail: xfliu@yzu.edu.cn
作者简介:段志强(1985-)，男，湖北襄阳人，讲师，博士，主要从事家禽重要疫病发病机制和免疫机制方面的研究，E-mail: zqduan@gzu.edu.cn
基金资助:
国家自然科学基金项目(31502074)；教育部“促进与美大地区科研合作与高层次人才培养项目”(教外司美［2014］2029号)；贵州省省校合作计划项目(黔科合LH字［2014］7669号)；公益性行业(农业)科研专项(201303033)

Genetic Variation and Pathogenicity Analysis of Virulent Newcastle Disease Viruses Isolated from Ducks in Guizhou Province

DUAN Zhi-qiang ^1，2，XU Hou-qiang^1，2，JI Xin-qin ^1，2，CHEN Qiang¹，HU Yan ¹，HU Shun-lin³，LIU Xiu-fan^3*

(1.College of Animal Science，Guizhou University，Guiyang 550025，China；2.Key Laboratory of
Animal Genetics，Breeding and Reproduction in the Plateau Mountainous Region of Ministry of
Education，Guizhou University，Guiyang 550025，China；3.Key Laboratory of Animal Infectious
Diseases of Ministry of Agriculture，Yangzhou University，Yangzhou 225009，China)

Received:2016-03-23 Online:2016-08-23 Published:2016-09-08

摘要/Abstract

摘要：

为了科学理解家鸭在新城疫病毒（NDV）流行和传播中的作用，对贵州不同地区分离的5株鸭源NDV强毒株进行遗传变异分析以及对鸭和SPF鸡的致病性研究。结果显示：5株NDV分离株均属于基因Ⅶd亚型，其F蛋白裂解位点基序均为112RRQKRF117，符合强毒株序列特征，与病毒致病性指数测定结果相符；F和HN蛋白中功能性氨基酸位点均高度保守，但在HN蛋白线性表位区有3株发生E347K突变。交叉血凝抑制试验发现分离株与传统疫苗株LaSota的抗原同源性较低（为83.3%~87.0%），而与新型基因Ⅶ型疫苗株A-Ⅶ的抗原同源性较高（为93.5%~100%）。将5株NDV分离株以0.5 mL病毒尿囊液原液通过肌肉注射感染鸭后未见明显发病死亡和病理变化，并且除脾外在其他多个组织脏器未能检测到病毒复制；而以10 ^6.0 ELD50•0.1 mL^－1剂量通过滴鼻点眼感染SPF鸡后6 d内100%发病死亡，病死鸡表现出新城疫典型的临床症状和病理变化，病毒在多个组织脏器中均能复制且对脾、胸腺和法氏囊等免疫器官损伤严重。另外，SPF鸡攻毒后喉头和泄殖腔棉拭病毒分离率明显高于试验鸭。本研究结果表明，贵州省鸭群中流行的基因Ⅶd亚型NDV强毒株HN蛋白发生E347K突变的变异株呈上升趋势，并与传统疫苗株产生抗原差异；5株鸭源NDV强毒株对鸭和鸡致病性差异显著，虽然对鸭无明显致病性，但鸭感染后可在较长时间内通过喉头或泄殖腔向体外排毒，因此必须采取有效措施防止NDV强毒由鸭群向鸡群的传播。

关键词: 新城疫病毒, 基因Ⅶd亚型, 遗传变异, 致病性

Abstract:

To better evaluate the role of domestic ducks in the epidemiology and transmission of NDV，five virulent NDV strains isolated from ducks in Guizhou Province were used to study their genetic variation and pathogenicity in ducks and SPF chickens.Sequence analysis showed that all the strains belonged to subgenotype Ⅶd NDV and had the velogenic motif 112RRQKRF117 in the cleavage sites of F protein，which was consistent with the results of biological tests.The functional amino acid sites in the F and HN proteins were all highly conserved，but three strains with E347K mutation were detected in the linear epitope of HN protein.Cross hemagglutination inhibition test revealed that the antigen homology of the isolates between LaSota and A-Ⅶ were 83.3%-87.0% and 93.5%-100%，respectively.In the challenge test，ducks of different groups challenged with virus allantoic fluid intramuscularly at the dose of 0.5 mL showed no obvious clinical symptoms and histopathologic changes，and viruses could not be detected in multiple tissues except for the spleen.On the contrary，SPF chickens infected with NDV strains intranasally at a dose of 10 ^6.0 ELD50•0.1 mL^－1 died in 6 days and showed typical clinical symptoms.Viruses could replicate in multiple tissues and cause severe damages to spleen，thymus and bursa in chickens.In addition，viruses were detected more frequently in the laryngeal and cloacal swabs of chickens than ducks.These results showed that subgenotype Ⅶd NDV strains with E347K mutation on HN protein significantly increased and had the antigenic differences with traditional vaccine strain.Although all the NDV strains were virulent in chickens whereas had no obvious pathogenicity in ducks，some measures should be taken to prevent virus transmission from ducks to chickens as viruses were detected in swabs of virulent NDV infected ducks for a long time.

Key words: Newcastle disease virus, subgenotype Ⅶd, genetic variation, pathogenicity

中图分类号:

段志强，许厚强，嵇辛勤，陈强，胡炎，胡顺林，刘秀梵. 贵州省鸭源新城疫病毒强毒株的遗传变异分析及致病性研究[J]. 畜牧兽医学报, doi: doi: 10.11843/j.issn.0366-6964.2016.08.013.

DUAN Zhi-qiang，XU Hou-qiang，JI Xin-qin，CHEN Qiang，HU Yan，HU Shun-lin，LIU Xiu-fan. Genetic Variation and Pathogenicity Analysis of Virulent Newcastle Disease Viruses Isolated from Ducks in Guizhou Province[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, doi: doi: 10.11843/j.issn.0366-6964.2016.08.013.

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贵州省鸭源新城疫病毒强毒株的遗传变异分析及致病性研究

Genetic Variation and Pathogenicity Analysis of Virulent Newcastle Disease Viruses Isolated from Ducks in Guizhou Province

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