| 鸡痘病毒FJ01株的分离鉴定 |
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| 引用本文： | 高映雪,林敏,胡换仪,刘小兰,刘昌锦,万文忠,罗锋,邓舜洲.鸡痘病毒FJ01株的分离鉴定[J].中国畜牧兽医,2020,47(2):575-582. |
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| 作者姓名： | 高映雪 林敏 胡换仪 刘小兰 刘昌锦 万文忠 罗锋 邓舜洲 |
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| 作者单位： | 1. 江西农业大学动物科学技术学院, 南昌 330045;2. 江西金伊博生物科技有限公司, 南昌 330013 |
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| 基金项目： | 国家自然科学基金项目（31460666）；江西省科技支撑计划项目（20132BBF60044、20141BBF60039）；江西省现代农业产业技术体系建设专项资金（JXARS-03） |
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| 摘 要： | 为调查福建规模化养鸡场病鸡的死亡原因，试验以福建省送检的鸡冠及头部皮肤有大量结节状痘痂的病鸡病料为研究对象进行了PCR鉴定。初步鉴定为鸡痘病毒（FWPV）株后，通过接种鸡胚绒毛尿囊膜（CAM）分离病毒；用原代鸡胚成纤维细胞（CEF）和传代细胞DF-1细胞对病毒进行传代，观察该分离毒株在两种细胞上培养特性的异同；对被感染的CEF进行超薄切片观察病毒的分布及病毒粒子的形态；并针对该分离株的TK基因和FPV175基因序列进行同源性分析。结果显示，接种经抗生素处理后的病料匀浆液上清的CAM出现大面积单个白色隆起的痘斑，匀浆痘斑后同时接种CEF和DF-1细胞，两种细胞均能产生稳定可持续传代的细胞病变，但病变出现的时间及病变程度不同；透射电镜下观察到典型的FWPV粒子密集分布在CEF的胞浆中，清晰可见卵圆形外膜包裹着的两侧凹陷的核心。对其中23个病毒粒子进行统计测得病毒粒子的大小为（258~344） nm×（153~238） nm；针对FWPV TK基因和FPV175基因的PCR检测及测序结果与GenBank收录的FWPV（登录号：NC_002188.1）核苷酸序列同源性分别高达100%和99.8%。以上结果表明该分离毒株为FWPV，命名为FWPV-FJ01，为国内FWPV的防治提供了参考依据。
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| 关 键 词： | 鸡痘病毒(FWPV) 分离鉴定 病毒粒子 福建分离株FJ01 |
| 收稿时间： | 2019-08-22 |
A Isolation and Identification of Fowlpox Virus FJ01 Strain |
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| GAO Yingxue,LIN Min,HU Huanyi,LIU Xiaolan,LIU Changjin,WAN Wenzhong,LUO Feng,DENG Shunzhou.A Isolation and Identification of Fowlpox Virus FJ01 Strain[J].China Animal Husbandry & Veterinary Medicine,2020,47(2):575-582. |
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| Authors: | GAO Yingxue LIN Min HU Huanyi LIU Xiaolan LIU Changjin WAN Wenzhong LUO Feng DENG Shunzhou |
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| Institution: | 1. College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang 330045, China;2. Jiangxi Jinyibo Biotechnology Company, Nanchang 330013, China |
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| Abstract: | To investigate the cause of death of diseased chickens in large-scale chicken farm in Fujian province,the diseased chickens with a large number of nodular scab in the crown and head skin were identified by PCR.After being identified as FWPV strain,the virus was isolated by inoculating chicken chorioallantoic membrane(CAM).The virus was subcultured with primary chicken embryo fibroblasts (CEF) and passage cells DF-1 cells,and the similarities and differences of the culture characteristics of the isolated strain on the two kinds of cells were observed.The distribution of the virus and the morphology of the virus particles were observed by ultra-thin sections of the CEF infected with the isolated strain.The homologies of TK and FPV175 genes were analyzed.The results showed that the CAM inoculated with antibiotics showed a large area of single white protuberant acne spots,and the CEF and DF-1 cells were inoculated at the same time.The two kinds of cells could produce stable and sustainable passage cytopathic effect,but the time and degree of the lesion were different.Under the transmission electron microscope,the typical fowlpox virus particles were densely distributed in the cytoplasm of the infected CEF,and the sunken cores wrapped by the oval outer membrane were clearly seen.According to the statistics of 23 virion particles,the size of virion particles was (258 to 344) nm & #215;(153 to 238) nm.PCR detection of FWPV TK gene and FPV175 gene showed that those nucleotide sequences of FWPV (accession No.:NC_002188.1) collected by GenBank were 100% and 99.8% homology,respectively.The above results indicated that the isolated strain was FWPV and was named as FWPV-FJ01,which provided a reference for the prevention and control of domestic FWPV. |
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| Keywords: | fowlpox virus (FWPV) isolation and identification virion Fujian isolate FJ01 |
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