4株NDV流行株的分离鉴定、生物学特性及遗传学特性的研究

doi:10.16431/j.cnki.1671-7236.2017.07.004

摘要/Abstract

摘要：

为研究近年国内的新城疫病毒（Newcastle disease virus，NDV）流行株与传统疫苗株（Mukteswar、La Sota及Clone 30）的亲缘关系和遗传演化情况，试验从广东、广西和海南地区患病的免疫鸡群病料中分离鉴定得到4株NDV流行株。通过RT-PCR技术对各毒株的F和HN基因进行扩增、克隆与序列分析，然后与GenBank上发表的NDV序列进行同源性比对、遗传进化树分析，并测定各毒株致病指数。结果显示，本试验中HN-08株的鸡胚半数感染量（EID₅₀）、平均死亡时间（MDT）、脑内致病指数（ICPI）和静脉致病指数（IVPI）分别为10^-8.37/0.2 mL、58.5 h、1.78和2.45，XX-08株分别为10^-6.50/0.2 mL、75.0 h、1.61和2.41，YS-09株分别为10^-7.75/0.2 mL、64.5 h、1.71和2.38，LF-09株分别为10^-7.60/0.2 mL、63.8 h、1.84和2.38。4株鸡源NDV流行株彼此间F和HN基因推导的氨基酸序列同源性在95.8%以上，与鸡源流行株GX11/03、GM株的同源性为96.8%~98.6%，而疫苗株Mukteswar、La Sota及Clone 30株的同源性为86.7%~90.6%，与标准强毒株F₄₈E₉的同源性为88.4%~91.3%。表明4株NDV流行株均为强毒株，与近年来国内流行的GX11/03、GM株有较近的亲缘关系，而与传统疫苗株的关系相对较远。

关键词: 新城疫病毒; F基因; HN基因; 遗传进化分析

Abstract:

In order to study the relationship and evolution between the latest Newcastle disease virus (NDV) epidemic strain and the vaccine strain (Mukteswar,La Sota and Clone 30 strains),four NDV epidemic strains were initially isolated and identified from immunized chicken groups in Guangdong,Guangxi and Hainan provinces. Fusion (F) gene and hemaglutinin neuraminidase (HN) gene of four isolated stains were amplified,cloned and sequenced by RT-PCR method,and the homology analysis of amino acid sequences deduced by F and HN genes of NDV and other strains published in GenBank were studied, the phylogenetic tree were build,and the pathogenicity index of each strain (EID₅₀,MDT, ICPI and IVPI) were determined. The results showed that the EID₅₀,MDT,ICPI and IVPI of the HN-08 strain were 10^-8.37/0.2 mL,58.5 h,1.78 and 2.45,that of the XX-08 strain were 10^-6.50/0.2 mL,75.0 h,1.61 and 2.41,that of the YS-09 strain were 10^-7.75/0.2 mL,64.5 h,1.71 and 2.38,and of the LF-09 strain were 10^-7.60/0.2 mL,63.8 h,1.84 and 2.38.In this experiment,the similarity of amino acid sequences of F and HN genes was over 95.8% among four strains of NDV isolated from chicken,the similarity of four NDV strains with chicken epidemic strains (GX11/03 and GM strains),vaccine strains (Mukteswar,La Sota and Clone 30 strains) and standard virulent strain (F₄₈E₉ strain) were from 96.8% to 98.6%,86.7% to 90.6% and 88.4% to 91.3%,respectively. The results showed that four NDV epidemic strains were virulent strains,which were closely related to the GX11/03 and GM strains that were popular in China in recent years,but relatively far from the traditional vaccine strains.

Key words: Newcastle disease virus; fusion (F) gene; hemaglutinin neuraminidase (HN) gene; phylogenetic analysis

中图分类号:

S855.3

李晓文, 刘佑明, 梁国智, 梅敏敏, 黄雯晶, 李文锋, 黄淑坚, 李智丽. 4株NDV流行株的分离鉴定、生物学特性及遗传学特性的研究[J]. 《中国畜牧兽医》, 2017, 44(7): 1925-1933.

LI Xiao-wen, LIU You-ming, LIANG Guo-zhi, MEI Min-min, HUANG Wen-jing, LI Wen-feng, HUANG Shu-jian, LI Zhi-li. Isolation, Identification of Four Newcastle Disease Virus Strains and Study on Their Biological Characteristics and Genetic Characteristics[J]. , 2017, 44(7): 1925-1933.

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