上海株犬冠状病毒的分离鉴定及S基因的遗传进化分析
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S852.65

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山东省潍坊市科学技术发展计划(2015GX054)


Isolation and Identification of Canine Coronavirus Shanghai Strain and Phylogenetic Analysis of S Gene
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    摘要:

    利用A72细胞(犬纤维肉瘤细胞系)从上海市某宠物医院临床表现为急性胃肠炎症状的幼犬腹泻粪便中分离到1株病毒,通过噬斑纯化、病毒理化特性检测以及动物回归实验和RT-PCR鉴定,证明所分离到的病毒为犬冠状病毒(Canine coronavirus,CCV),将其命名为CCV-SHdc株。根据GenBank公布的CCV S基因序列,设计合成4对特异性引物,利用RT-PCR成功扩增获得CCV-SHdc株S基因序列,同时进行序列测定和核苷酸系统发育进化树绘制。结果显示,CCV-SHdc株的S基因序列全长4 364 bp;该毒株与日本分离的CCV 5821株亲缘关系最近,处于同一分支,与意大利分离的CCV 23/03和CCV Elmo/02株亲缘关系较远。核苷酸与氨基酸同源性分析表明,CCV-SHdc株与CCV 23/03和CCV Elmo/02株可能属于不同基因型,已有很多点发生突变,这种突变的累积有可能引起病毒毒力的变化。

    Abstract:

    A strain of virus was isolated by A72 cells from feces of a diarrhea puppy with clinical manifestation of acute gastroenteritis in a pet clinic in Shanghai city. Through identification experiments containing plaque purification,detection of physicochemical properties,animal regression test,and RT-PCR method,the virus strain was confirmed as canine coronavirus(CCV)and named CCV-SHdc. Based on the sequence of S gene of CCV published in GenBank,4 pairs of primers were designed to amplified S gene by RT-PCR. Then sequencing the amplified gene and drawing a phylogenetic tree. Results indicated that the length of S gene of CCV-SHdc was 4 364 bp. It was closest to strain CCV 5821 that isolated from Japan,belonging to the same one clade,however,it has a farther relationship with CCV 23/03 and CCV Elmo/02 of Italy strain. Analysis of amino acid homology and nucleotide sequence homology indicated genotype of CCV-SHdc strain may be different from strains of CCV 23/03 and CCV Elmo/02. CCV-SHdc strain had multipoint mutation,the accumulation of which would lead to virulence change

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沈美艳,孙秋艳,李舫,王彩霞.上海株犬冠状病毒的分离鉴定及S基因的遗传进化分析[J].《中国动物检疫》编辑部,2017,34(3):97-101.

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  • 在线发布日期: 2017-03-07
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