用鸡胚培养对虾白斑综合症病毒(WSSV)的研究
PDF下载 (303)刘飞,李登峰*.用鸡胚培养对虾白斑综合症病毒(WSSV)的研究[J].宁波大学学报(理工版),2017,30(1):47-51.DOI:
LIU Fei,LI Deng-feng*.Propagation of white spot syndrome virus (WSSV) in embryonated eggs[J].Journal of Ningbo University(Natural Science & Engineering Edition),2017,30(1):47-51.DOI:
| Title: | Propagation of white spot syndrome virus (WSSV) in embryonated eggs |
| 作者: | 刘飞, 李登峰* |
| Author(s): | LIU Fei, LI Deng-feng* |
| 关键词: | 对虾白斑综合症病毒; 鸡胚; 培养 |
| Keywords: | white spot syndrome virus; chicken embryonated eggs; cultivation |
| 分类号: | — |
| 文献标识码: | A |
| 摘要: | 为探索对虾白斑综合症病毒(WSSV)能否在鸡胚中培养, 基于稳定培养体系, 将WSSV接种到SPF鸡胚中. 收获接种3d后死亡的鸡胚, 从中提取病毒; 将其再次接种到健康鸡胚中, 传代5次后, 由卵黄囊膜提取病毒, 将其经肌肉注射至克氏原螯虾, 结果实验组克氏原螯虾100%死亡, 而对照组无死亡情况. 对各代鸡胚及克氏原螯虾进行PCR和电镜检测, 实验组呈WSSV阳性; 电镜下, 死亡鸡胚细胞中可见大量正在组装的病毒粒子; 表明WSSV可用鸡胚进行培养. |
| Abstract: | In order to establish a stable system of cultivating white spot syndrome virus (WSSV), the specific pathogen free chicken embryonated eggs are inoculated with WSSV suspensions. Both the yolk sac membrane and yolk sac fluid are harvested to prepare passage inoculation suspensions. Passages 2-5 are carried out by inoculating SPF CEE via the yolk sac route. Yolk sac membrane and yolk sac fluid are detected via transmission electron microscope (TEM) observation and PCR. A great deal of WSSV particles are identified in the ultra-thin sections of experimental groups. Positive PCR results in experimental groups inoculated with WSSV and negative results in control groups are obtained. In artificial infection, WSSV isolated from the fifth passage of experimental chicken embryo results in 100% mortality to Procambarus clarkia, which confirms the success of replication of WSSV in CEE |
| 参考文献 /References: | [1].ESCOBEDO-BONILLA C M, ALDAY-SANZ V, WILLE M, et al. A review on the morphology, molecular characterization, morphogenesis and pathogenesis of white spot syndrome virus [J]. Journal of Fish Diseases, 2008, 31(1):1-18. [2].王清印, 杨丛海. 中国对虾健康养殖的发展现状及展望[J]. 中国水产, 2005, (1):21-24. [3].LEU J H, YANG F, ZHANG X, et al. Whispovirus[J]. Current Topics in Microbiology and Immunology, 2009, 328:197-227. [4].DIEU B T M, MARKS H, ZWART M P, et al. Evaluation of white spot syndrome virus variable DNA loci as molecular markers of virus spread at intermediate spatiotemporal scales[J]. Journal of General Virology, 2010, 91:1164-1172. [5].REN Q, HUANG Y, HE Y D, et al. A white spot syndrome virus microRNA promotes the virus infection by targeting the host STAT[J]. Scientific Reports, 2015, 5:1-12. [6].LI Z, LI F, HAN Y, et al. VP24 is a chitin-binding protein involved in white spot syndrome virus infection [J]. Journal of Virology, 2015, 90(2):842-850. [7].PAZIR M K, AFSHARNASAB M, NIAMAYMANDI N, et al. Histopathological observation of white spot syn- drome virus and infectious hypodermal and hemato- poiectic necrosis virus in shrimp farms, Litopenaeus vannamei, in Bushehr Province, Iran[J]. Asian-Australa- sian Journal of Animal Sciences, 2012, 6(5):209-219. [8].朱建中, 陆承平. 对虾白斑综合症病毒在螫虾动物模型的感染特性[J]. 水产学报, 2001, 25(1):47-51. [9].徐丽美, 王玮, 杨丰. 对虾白斑杆状病毒(WSSV)定量PCR检测技术研究[J]. 高技术通讯, 2001, 1):41-61. [10].胡广洲. 中华鳖暴发性败血症的研究[D]. 宁波: 宁波大学, 2010. [11].张春杰, 程相朝. 不同途径接种鸡胚培养传染性法氏囊病病毒的比较研究[J]. 中国兽医科技, 1996, 26(4): 25-27. [12].GUY J S. Isolation and propagation of coronaviruses in embryonated eggs[J]. Methods in Molecular Biology, 2015, 1282:63-71. [13].NU?EZ L F, PARRA S H, METTIFOGO E, et al. Isolation of chicken astrovirus from specific pathogen- free chicken embryonated eggs[J]. Poultry Science, 2015, 94(5):947-954. [14].姜有声, 邢, 王世表等. 白斑综合症病毒的体外增殖研究[J]. 水产学报, 2004, 28:93-96. [15].XIE X X, LI H Y, XU L M, et al. A simple and efficient method for purification of inact white spot syndrome virus (WSSV) viral particles[J]. Virus Research, 2005, 108:63-67. [16].中国医学科学院流行病防治研究所. 常见病毒实验技术[M]. 北京科学出版社, 1978:25-32. [17].AL-MUBARAK F, DALY J, CHRISTIE D, et al. Identification of morphological differences between avian influenza A viruses grown in chicken and duck cells[J]. Virus Research, 2015, 199:9-19. [18].HSUEH T Y, GIBSON K E. Transfer of Acanthamoeba spp. to fresh produce from water and environmental surfaces[J]. Letters in Applied Microbiology, 2015, 61(2): 192-198. [19].LASCOLA B. Looking at protists as a source of patho- genic viruses[J]. Microbial Pathogenesis, 2014, 77:131- 135. |
| 备注/Memo: | 收稿日期: 2016?01?10. 宁波大学学报（理工版）网址: http://journallg.nbu.edu.cn/ 基金项目: 国家科技部星火计划（2014GA701018）; 宁波市科技局农业攻关项目（2011C10004）; 浙江省教育厅项目（Y201327815）; 浙江省重点学科开放基金（E02009124200）; 宁波大学科研基金（XKZSC1519）; 宁波大学王宽诚幸福基金. 第一作者: 刘飞（1991－）, 女, 安徽滁州人, 在读硕士研究生, 主要研究方向: 水生生物病毒. E-mail: lvphy0708@126.com *通信作者: 李登峰（1975－）, 女, 湖南双峰人, 副教授, 主要研究方向: 水产动物病害防治. E-mail: lidengfeng@nbu.edu.cn 宁波大学学报（理工版）网址：http://journallg.nbu.edu.cn/ |
