| 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，8 000 r/min离心15 min取上清，接种10日龄鹅胚的卵黄囊，收集24~144 h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380 bp，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。 |
| 关键词: 鹅呼肠孤病毒 分离鉴定 致病性 |
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| 基金项目: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） |
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| Isolation,identification and pathogenicity of a goose reovirus strain |
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ZHANG Bingqian,DIAO Youxiang,YU Xianglong,et al
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| Abstract: |
| 【Objective】A strain of goose reovirus was isolated and identified from suspected infectious materials and its pathogenicity was evaluated.【Method】The liver and spleen tissues of dead geese were homogenized and frozen for three times.Supernatant was obtained after 8 000 r/min centrifugation for 15 minutes.A virus was isolated from allantoic fluid of 10 days old goose embryo by yolk sac inoculation.Specific primers were designed according to the conserved sequence of goose reovirus gene,and RT PCR was conducted to the extracted RNA.The band was recovered and cloned into pMD18-T vector,and the recombinant vector was transformed into Escherichia coli DH5α and cultured in AMP/LB medium.The sequence of positive colonies was determined and the nucleotide and amino acid homology of measured sequence was conducted to construct phylogenetic tree.【Result】The length of the isolated JS-01 sequence was 380 bp,and it was a fragment of goose reovirus sigmaC gene.The isolate and waterfowl reovirus belonged to same branch with the homology of 94.8%-96.5%,while the homology with chicken reovirus was only 40.6%-52.2%.Animal regression experiment showed that JS 01 could cause liver,spleen,kidney and other organ lesions to goslings.【Conclusion】The isolated strain was goose reovirus and had pathogenicity to gosling.Attention should be paid to the prevention and control of the disease. |
| Key words: goose reovirus isolation and identification pathogenicity |
