鸡传染性支气管炎病毒HN_(99)株的分离与鉴定  

Isolation and identification of an avian infectious bronchitis virus strain HN99

作　　者：高文明[1] 崔保安[1] 李新生[1] 陈红英[1] 王学斌[1] 吕晓丽[1] 

机构地区：[1]河南农业大学牧医工程学院,河南郑州450002

出　　处：《西北农林科技大学学报（自然科学版）》2008年第4期47-52,共6页Journal of Northwest A&F University(Natural Science Edition)

基　　金：河南省"十一五"重大科技专项(0620010300)

摘　　要：【目的】分离鸡传染性支气管炎病毒河南地方株,并对其进行鉴定。【方法】采用鸡胚盲传,观察病毒对鸡胚的致病作用；通过酶处理、病毒干扰试验、动物回归试验、理化试验等研究病毒的特性；利用电镜观察病毒的形态,应用RT-PCR方法扩增分离毒株的S1和N基因片段。【结果】病毒盲传至第2代后鸡胚出现死亡和侏儒胚,将病毒盲传至第8代时,病毒的毒力逐渐由10^4.0增强到10^7.2；经质量分数1%胰酶处理或用磷酸酯酶C处理的病毒液能够明显凝集质量分数1%鸡红细胞,血凝滴度分别在2^9和2^7左右；病毒能够干扰NDV在鸡胚中的增殖,单独接种分离毒株的尿囊液HA滴度平均为2^0,先接种HN99毒株再接种NDV La Sota株的尿囊液HA滴度平均为2^3,同时接种HN99毒株和NDV La Sota株的尿囊液HA滴度平均为28.5,单独接种NDV La Sota株的尿囊液HA滴度平均为2^9；病毒可致SPF雏鸡发病,引起肾脏肿大、花斑肾、尿酸盐沉积等肾脏病变；该毒株对乙醚和氯仿敏感,耐酸不耐碱,对热敏感；病毒粒子直径90-105 nm；扩增到了分离毒株S1基因片段大小为1 739 bp（含前导序列）,N基因全长为1 230 bp,通过与其他IBV毒株进行系统进化分析,证明分离毒株与其他毒株的亲缘关系较远。【结论】初步证实分离的病毒为肾型鸡传染性支气管炎病毒,命名为HN99株。[Objective] An experiment was carried out to isolate infectious bronchitis virus （IBV） from the diseased chickens in Henan Province. [Method] Biological characteristics of the virus were studied by virus pathogenicity to chicken embryo, hemagglutination（HA） activity and interference effect on replication of NDV,pathogenicity to SPF chickens, physiochemical characteristics of the virus were also studied and morphological observed by electron-microscope. S1 and N genes were amplified by reverse transcription-polymerase chain reaction （RT-PCR）. [Result] The second passage virus produced gross lesions to chicken embryos. The virus could hemagglutinate chicken red blood cells about 29 and 27 after treatment with enzyme or phosphoesterase. Replication of NDV was affected by inoculating in the same chicken embryo after inoculation with the isolated virus. The morbidity and mortality of the four passage virus to 7-day-old chickens were 900/40 and 400/40. Moreover,the gross lesions in kidney caused by the virus were distinct. In addition, physiochemical properties of the virus shared the typical characteristics of most strains of IBV. The virus showed typical shape shared by members of Coronaviridae under electron-microscope. S1 and N genes were amplified by RT-PCR and sequenced by automatic sequencing system. S1 gene consisted of 1 739 bp, N gene was 1 230 bp. Evolution analysis indicated that the isolated virus was distantly related to the other strains of IBV. [Conclusion] All the results showed that the virus was a new IBV variant in China,and it was tentatively named HN99.

关 键 词：鸡传染性支气管炎病毒 肾型 分离与鉴定 

分 类 号：S852.659.6[农业科学—基础兽医学]