一株鹅源呼肠孤病毒的分离鉴定及致病性研究

发布时间：2018-06-15 16:13

本文选题：鹅呼肠孤病毒 + 分离鉴定　；参考：《西北农林科技大学学报(自然科学版)》2017年08期

【摘要】：【目的】从疑似感染鹅呼肠孤病毒的病料中分离鉴定一株鹅源呼肠孤病毒JS-01,并对病毒分离株的致病性进行初步研究,为开展该病毒的进一步研究奠定基础。【方法】取病死鹅的肝、脾组织冻融3次后充分匀浆,8 000r/min离心15min取上清,接种10日龄鹅胚的卵黄囊,收集24~144h死亡的鹅胚尿囊液,并提取病毒RNA。根据鹅呼肠孤病毒基因的保守序列设计特异性引物,利用该引物对所提取的病毒RNA进行RT-PCR,回收后与pMD18-T载体连接,将重组载体转化到DH5α大肠杆菌并在AMP+/LB培养基中培养,对阳性菌落测序,对测得的序列进行核苷酸及氨基酸同源性比对,并绘制系统遗传进化树。【结果】分离病毒JS-01的RT-PCR显示,其基因序列长度380bp,为鹅呼肠孤病毒σC基因片段;系统遗传进化分析表明,分离株JS-01与水禽源呼肠孤病毒处于同一分支,同源性为94.8%~96.5%,而与鸡呼肠孤病毒的同源性仅为40.6%~52.2%;在雏鹅上进行的动物回归试验表明,病毒JS-01可使雏鹅肝、脾、肾等器官发生病变。【结论】分离毒株为鹅呼肠孤病毒,该分离株对雏鹅有一定致病性,应注意本病的防控。
[Abstract]:[objective] to isolate and identify a geese origin reovirus JS-01 from the samples suspected to be infected with geese reovirus, and to study the pathogenicity of the isolated strain. [methods] the liver and spleen tissue of the dead goose were taken for 3 times, and then fully homogenized for 8 000r/min centrifuged 15min supernatant. The egg yolk sac of 10 day old goose embryo was inoculated, and the allantoic fluid of the dead goose embryo was collected. And the virus RNA was extracted. According to the conserved sequence of geese reovirus gene, a specific primer was designed, using the primer to carry out RT-PCR on the extracted virus RNA, and the recombinant vector was ligated with pMD18-T vector after recovery. The recombinant vector was transformed into DH5 伪 Escherichia coli and cultured in AMP / LB medium. The nucleotide and amino acid homology of the positive bacteria was sequenced, and the phylogenetic tree was plotted. RT-PCR of JS-01 virus showed that the length of the gene was 380 BP, which was a 蟽 C gene fragment of geese reovirus. The phylogenetic analysis showed that the isolated strain JS-01 was in the same branch as the waterfowl origin reovirus, and the homology was 94.88.96. 5, while the homology with chicken reovirus was only 40.6 and 52.2, and the animal regression test on the goslings showed that the virus JS-01 could make foie liver. [conclusion] the isolated strain is goose reovirus, which is pathogenicity to goslings, so we should pay attention to the prevention and control of this disease.
【作者单位】：山东农业大学动物科技学院;
【基金】：现代农业产业技术体系建设专项(CARS-43-10) 山东省科技发展计划项目(2014GNC111023)
【分类号】：S858.33

【相似文献】