山羊Aichivirus C的分离鉴定及演化分析OA北大核心CSTPCD
Isolation and Molecular Characterization of Aichivirus C from Goats
C型爱知病毒(Aichivirus C)是新近证实的致山羊腹泻病毒,本研究旨在分离山羊Aichivirus C并分析其基因组特征.在四川某暴发腹泻的山羊场采集腹泻粪便9份,1只病死羔羊的组织样本13份,进行常见的腹泻病原检测.采用Vero细胞系对山羊Aichivirus C阳性样本进行病毒分离,以PCR、间接免疫荧光试验和电镜技术对分离毒株进行鉴定,并扩增获得其基因组序列.结果显示:通过RT-PCR方法从9份山羊腹泻粪便样本中检出Aichivirus C阳性8份,羔羊的组织样本中检出Aichivirus C阳性8份,其它常见的致山羊腹泻病原在上述粪便和组织样本中均未检出.使用Vero细胞系成功从阳性样品中分离到2株山羊Aichivirus C,其病毒滴度分别为106.5 TCID50·mL-1和105.9 TCID50·mL-1.获得两条近乎全长基因组序列,与GenBank中山羊源Aichivirus C毒株的核苷酸相似性为78.8％～97.4％,与国内SWUN/F11/2019毒株的核苷酸相似性为97.0％～97.4％.此外,本研究还从5份临床阳性样本中扩增出VP0、VP3和VP1基因完整序列.基于基因组的演化分析显示:获得的两个病毒株与SWUN/F11/2019毒株在系统发育树上聚为一支.并且,这些毒株的VP0和VP1基因编码蛋白也有独特的氨基酸突变.综上所述,本研究分离得到2株山羊Aichivirus C,获得了其近乎全长基因组序列,与国内SWUN/F11/2019毒株亲缘关系最近,其VP0和VP1基因具有独特的分子特征.本结果丰富了国内山羊源Aichivirus C的分子遗传信息,为进一步研究山羊Aichivirus C的致病性和分子生物学特征提供了参考.
Aichivirus C is a newly confirmed virus associated with caprine diarrhoea.The purpose of this study was to isolate goat Aichivirus C and analyze the genomic characteristics.Nine fecal samples and 13 tissue samples from a dead kid were collected from a goat farm with an outbreak of diarrhoea in Sichuan province,and used for testing the common pathogens associated with di-arrhoea.Virus was isolated from Aichivirus C positive samples using Vero cells and identified by PCR,indirect immunofluorescence and electron microscopy.The complete genomes of the iso-lates were amplified and analyzed.Results were as follows:8 fecal samples with diarrhoea and 8 tissue samples from a lamb were detected as positive for Aichivirus C by RT-PCR,and no other common diarrhoea-causing pathogens were detected in the above samples.Two Aichivirus C strains were successfully isolated from positive samples using Vero cells,and the virus titers were 106.5TCID50·mL-1 and 105.9TCID50·mL-1,respectively.The nearly full-length genomic se-quences of both strains were obtained,which shared 78.8％-97.4％nucleotide identity with the known goat Aichivirus C genomes from GenBank and 97.0％-97.4％nucleotide identity with the SWUN/F11/2019 isolate from China.Furthermore,the VP0,VP3 and VP1 genes were success-fully amplified from 5 clinical positive samples.Phylogenetic analysis based on the genome re-vealed that the two isolates were clustered with the SWUN/F11/2019 strain.Unique amino acid mutations were also revealed in the VP0 and VP1 genes.In summary,two strains of goat Aichi-virus C were successfully isolated and the nearly full-length genomic sequences were obtained.Phylogentic analysis demonstrated that these two strains are closely related to the SWUN/F11/2019 strain from China,with a unique molecular characteristics in the VP0 and VP1 genes.The results enrich the molecular genetic information of Aichivirus C from goats in China and laid the foundation for further studies on the pathogenicity and molecular biological characteristics of goat Aichivirus C.
程玉婷;阿比克哈莫;杨晨;张丁中;任云鑫;岳华;汤承
西南民族大学畜牧兽医学院,成都 610041
畜牧业
山羊腹泻C型爱知病毒分离鉴定分子特征
goatdiarrheaAichivirus Cisolationmolecular characterisation
《畜牧兽医学报》 2024 (008)
3612-3622 / 11
西南民族大学"双一流"项目资助(XM2023014)
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