| 猪δ冠状病毒的分离鉴定及其遗传进化分析 |
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| 引用本文： | 方谱县,刘康,张蕙畅,夏思进,张健淞,任杰,肖少波,方六荣.猪δ冠状病毒的分离鉴定及其遗传进化分析[J].华中农业大学学报,2021,40(6):168-175. |
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| 作者姓名： | 方谱县 刘康 张蕙畅 夏思进 张健淞 任杰 肖少波 方六荣 |
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| 作者单位： | 华中农业大学农业微生物学国家重点实验室/华中农业大学动物医学院/
湖北省生猪健康养殖协同创新中心，武汉 430070 |
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| 基金项目： | 国家自然科学基金项目（32072846）；国家自然科学基金青年科学基金项目（31902247） |
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| 摘 要： | 通过RT-PCR对采集自河北省2个猪场的5份腹泻仔猪小肠内容物进行检测，结果均为猪δ冠状病毒（PDCoV）阳性。将病料处理后接种LLC-PK1细胞，从2个猪场的病料中各成功分离到1株PDCoV，分别命名为PDCoV CHN-HeB-A1株和CHN-HeB-B2株。通过间接免疫荧光试验（IFA）、Western blot和电镜观察对分离的病毒进一步鉴定，IFA和Western blot结果证实2株病毒都能与PDCoV M蛋白的单克隆抗体发生特异性反应，电镜下可观察到直径100～150 nm、呈典型皇冠状的病毒粒子，证实所分离的病毒为PDCoV。全基因组测序以及遗传进化分析结果表明，2株PDCoV与中国大陆毒株相似性较高，为98.7％~99.1％，与中国香港毒株处于同一簇。
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| 关 键 词： | 猪δ冠状病毒 分离 鉴定 基因组测序 遗传进化 |
| 收稿时间： | 2021/5/28 0:00:00 |
Isolation,identification and genetic evolution analysis of porcine deltacoronavirus |
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| FANG Puxian,LIU Kang,ZHANG Huichang,XIA Sijin,ZHANG Jiansong,REN Jie,XIAO Shaobo,FANG Liurong.Isolation,identification and genetic evolution analysis of porcine deltacoronavirus[J].Journal of Huazhong Agricultural University,2021,40(6):168-175. |
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| Authors: | FANG Puxian LIU Kang ZHANG Huichang XIA Sijin ZHANG Jiansong REN Jie XIAO Shaobo FANG Liurong |
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| Abstract: | Porcine deltacoronavirus (PDCoV) is a newly emerged enteric coronavirus. In this study,five samples of piglet intestinal contents with diarrhea from two pig farms in Hebei Province were identified to be positive for PDCoV by RT-PCR assay,followed by the virus isolation through inoculation of treated samples on LLC-PK1 cells. The result showed that two PDCoV strains from various pig farms were successfully isolated and named as PDCoV CHN-HeB-A1 and CHN-HeB-B2 strains respectively,which were further identified by indirect immunofluorescence test (IFA),Western blot and electron microscopy assay. The results of IFA and Western blot confirmed that both virus strains can specifically react with monoclonal antibodies against PDCoV M protein. Virus particles with a diameter of 100-150 nm and a typical crown shape were obviously observed under electron microscopy,verifying both virus strains to be PDCoV. The results of whole genome sequencing and genetic evolution analysis showed that both PDCoV strains were homologous to the Chinese strain,with the similarity ranging from 98.7% to 99.1%,which were in the same cluster as the Hong Kong strain. |
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| Keywords: | porcine detalcoronavirus isolation identification genome sequencing genetic evolution |
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