1株基因Ⅶ型新城疫病毒的生物学特性研究及全基因组序列分析

doi:10.16431/j.cnki.1671-7236.2018.10.021

摘要/Abstract

摘要：

为研究当前流行的新城疫病毒（Newcastle disease virus，NDV）基因型、致病性及其与传统新城疫病毒疫苗株（La Sota等）的核苷酸差异，试验从某发病鸡场病死鸡体内分离到1株疑似NDV毒株，经红细胞凝集试验（HA）和红细胞凝集抑制试验（HI）初步确定为鸡源NDV。参照GenBank公布的NDV F基因部分片段（登录号：JF950510.1）设计1对引物，通过RT-PCR技术扩增分离株的F基因并克隆、测序，测序结果与NCBI中NDV F基因序列进行比对，构建系统进化树并分析其基因型；通过测定鸡胚平均致死时间（MDT）、1日龄鸡脑内致病指数（ICPI）和6周龄鸡静脉致病指数（IVPI）判断病毒致病性；参照GenBank公布的NDV全基因组序列（登录号：JF950510.1）设计9对引物对分离株进行全基因组序列测定，并分析其基因组结构。结果表明，RT-PCR扩增得到F基因长约500 bp，基于F基因构建的系统进化树显示分离株为基因Ⅶ型NDV；MDT、ICPI和IVPI分别为52.8 h、1.675和2.46，表明分离株属于强毒株。全基因组序列分析显示，分离株全基因组全长15 192 bp，与传统La Sota株基因组相比，序列多出6个碱基，核苷酸序列同源性82.8%。本研究成功分离到1株基因Ⅶ型NDV强毒株，且与传统疫苗毒株La Sota的核苷酸序列同源性差异较大。

关键词: 新城疫病毒(NDV); F基因; 强毒株; 全基因组

Abstract:

In order to study the genotype,pathogenicity and nucleotide difference between Newcastle disease virus (NDV) isolates and the traditional NDV vaccine strain (La Sota),a suspected NDV virus was isolated from a chicken farm.The isolate was preliminarily determined by HA and HI tests.A pair of primers was designed based on the partial fragment of NDV F gene published at GenBank(accession No.:JF950510.1).F gene was amplified by RT-PCR method,and cloned and sequenced,the sequencing result was compared with the F gene sequences published at GenBank,and the evolutionary tree was constracted to analyze the genotypes.The pathogenicity of the virus was determined by the mean death time (MDT) of chicken embryos,intracerebral pathogenicity index (ICPI) in one-day-old chickens and intravenous pathogenicity index (IVPI) in six-week-old chickens,respectively.Based on the NDV genome sequence published in GenBank(accession No.:JF950510.1),nine pairs of primers were designed to amplify the genome sequence of the isolate strain,and its structure was analyzed.The results showed that the length of F gene was about 500 bp,a NDV strain of genotype Ⅶ was isolated.The MDT,ICPI and IVPI were 52.8 h,1.675 and 2.46,respectively.The whole genome sequence analysis results showed that the full genome length of the isolated strain was 15 192 bp,and had 6 nucleotides more than the strain of La Sota,and the homology between the two strains was 82.8%.Therefore,a virulent NDV strain of genotype Ⅶ was isolated,and contained a large difference in the nucleotide sequence compared with the La Sota strain.

Key words: Newcastle disease virus(NDV); F gene; virulent strain; whole genome

中图分类号:

S852.65⁺9.5

孙化露, 毕云英, 于泽坤, 李思菲, 刘阳, 李明义, 崔现兰. 1株基因Ⅶ型新城疫病毒的生物学特性研究及全基因组序列分析[J]. 《中国畜牧兽医》, 2018, 45(10): 2840-2848.

SUN Hualu, BI Yunying, YU Zekun, LI Sifei, LIU Yang, LI Mingyi, CUI Xianlan. Biological Characterization and Complete Genomic Analysis of a Genotype Ⅶ Newcastle Disease Virus Strain[J]. , 2018, 45(10): 2840-2848.

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