1株赛鸽新城疫强毒株的致病特性及分子特征分析

doi:10.15933/j.cnki.1004-3268.2019.10.021

河南农业科学 ›› 2019, Vol. 48 ›› Issue (10): 140-146.DOI: 10.15933/j.cnki.1004-3268.2019.10.021

1株赛鸽新城疫强毒株的致病特性及分子特征分析

李珮瑶¹，高晶萍¹，田勇¹，王存连¹，徐明举¹，利凯¹，李军²，张瑞华¹，兰金苹²，徐彤^1,2

（1.河北北方学院预防兽医学重点实验室，河北张家 075000； 2.河北北方学院生命科学研究中心，河北张家口 075000）

收稿日期:2019-03-21 出版日期:2019-10-15 发布日期:2019-10-15
通讯作者: 徐彤（1969-），男，河北承德人，教授，博士，主要从事动物流感病毒诱导肺损伤机制研究。E-mail:xutong1969@sohu.com
作者简介:李珮瑶（1996-），女，河北邯郸人，在读硕士研究生，研究方向：动物疫病防控。E-mail：peiyaoli19@163.com
基金资助:
河北省第二期现代农业产业技术体系蛋肉鸡创新团队建设专项（HBCT2018150207）；河北省教育厅在读研究生创新能力资助项目(CXZZSS2018129)；赛鸽主要疫病防控技术体系研究项目（自选课题）

Pathogenicity Analysis and Molecular Characterization of Newcastle Disease Virus Isolated from Ｒacing Pigeon

LI Peiyao¹，GAO Jingping¹，TIAN Yong¹，WANG Cunlian¹，XU Mingju¹，LI Kai¹，LI Jun²，ZHANG Ｒuihua¹，LAN Jinping²，XU Tong¹

( 1.Key Laboratory of Preventive Veterinary Medicine,Hebei North University,Zhangjiakou 075000,China; 2.Life Science Ｒesearch Center,Hebei North University,Zhangjiakou 075000,China)

Received:2019-03-21 Published:2019-10-15 Online:2019-10-15

摘要/Abstract

摘要： 为了解赛鸽群中新城疫流行毒株的分子遗传特征及变异情况，从河北省疑似赛鸽新城疫病例中采集病料，常规处理后接种SPF鸡胚，经血凝试验（HA）和血凝抑制试验（HI）鉴定病毒，并对分离毒株进行致病性试验、毒力测定，采用RT-PCR方法扩增F基因，并与其他不同种属新城疫毒株进行遗传进化发育分析和氨基酸序列同源性分析。结果显示，从病料中分离到1株新城疫病毒（命名为HB株），致病性试验产生与发病鸽相似的临床症状和剖检变化；其鸡胚平均死亡时间（MDT）、1日龄雏鸡脑内致病指数（ICPI）和6周龄雏鸡静脉致病指数（IVPI）分别为52.8 h、1.78和2.59，表明HB株新城疫病毒为强毒株；HB株F基因碱基序列包含1个大小为1 662 bp的完整开放阅读框，基因分型结果显示，HB株属于基因型Ⅵ型。根据遗传进化发育关系和同源性分析结果发现，HB株与pi/CH/LGD/110208、dove/Italy/2736/00、Q-GB 506/97等毒株核苷酸序列相似性和氨基酸同源性较高，尤其与pigeon/Qinghai/1344/2017毒株亲缘关系最近，分别达98.45%和99.5%；与目前使用的传统新城疫疫苗毒株LaSota、B1、V4、Mukteswar株等遗传关系较远;F蛋白裂解位点氨基酸序列为¹¹²R-R-Q-K-R-F¹¹⁷，符合强毒株特征。以上结果表明，HB株为强毒株且与目前广泛使用的新城疫疫苗毒株遗传关系较远。

关键词: 赛鸽, 新城疫, F基因, 致病特性, 分子特征

Abstract: In order to understand the molecular genetic characteristics and variation of Newcastle disease epidemic strains in racing pigeon population,in this experiment,the disease material was collected from the suspected pigeon Newcastle disease case in Hebei Province.After routine treatment,the SPF chicken embryo was inoculated.The chicken embryo allantoic fluid was collected and identified by Hemagglutina tion and Hemagglutination inhibition test as Newcastle disease virus,and pathogenicity tests,virulence as says were conducted.The sequences of F gene were amplified by ＲT-PCＲ and genetic evolution and ami no acid sequence homology with other different species of Newcastle disease virus strains were analyzed． The results showed that a Newcastle disease virus(named HB strain) was isolated from the disease mate rial.The pathogenicity test of HB strain presented similar clinical symptoms and necropsy changes with those found in naturally diseased pigeons． The mean death time，intracerebral pathogenicity index and in travenous pathogenicity index of HB strain were 52.8 h,1.78,and 2.59，which indicated that the HB strain was a virulent strain.F gene of HB strain contained a complete open reading frame(OＲF) of 1 662 bp.The genotyping results showed that the HB strain belonged to the Ⅵ genotype.Genetic relationship of HB strain showed that it was closed to pi/CH/LGD/110208,dove/Italy/2736/00,Q-GB 506/97 based on F gene.Especially with the pigeon/Qinghai/1344/2017 strain，the closest relativities of nucleotide and amino acid sequences were 98.45% and 99.5%.However,it had far genetic relationship with traditional chicken Newcastle disease vaccine strains of B1,LaSota,V4 and Mukteswar.The F protein cleavage site amino acid sequence of HB strain was 112Ｒ-Ｒ-Q-K-Ｒ-F117,which was consistent with the characteristics of virulent strains.The results indicate that the HB strain is a virulent strain and has a large genetic differ ence from the current widely used Newcastle disease vaccine strains．

Key words: Ｒacing pigeon, Newcastle disease, F gene, Pathogenic characteristics, Molecular charac terizaion

中图分类号:

S855.3

李珮瑶, 高晶萍, 田勇, 王存连, 徐明举, 利凯, 李军, 张瑞华, 兰金苹, 徐彤. 1株赛鸽新城疫强毒株的致病特性及分子特征分析[J]. 河南农业科学, 2019, 48(10): 140-146.

LI Peiyao, GAO Jingping, TIAN Yong, WANG Cunlian, XU Mingju, LI Kai, LI Jun, ZHANG Ｒuihua, LAN Jinping, XU Tong. Pathogenicity Analysis and Molecular Characterization of Newcastle Disease Virus Isolated from Ｒacing Pigeon[J]. Journal of Henan Agricultural Sciences, 2019, 48(10): 140-146.

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1株赛鸽新城疫强毒株的致病特性及分子特征分析

Pathogenicity Analysis and Molecular Characterization of Newcastle Disease Virus Isolated from Ｒacing Pigeon

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