中国媒介生物学及控制杂志 ›› 2021, Vol. 32 ›› Issue (4): 412-414.DOI: 10.11853/j.issn.1003.8280.2021.04.005

• 媒介生物病原学监测专题 • 上一篇    下一篇

库蚊黄病毒的病毒滴度测定方法建立

何英, 殷启凯, 付士红, 王环宇   

  1. 中国疾病预防控制中心病毒病预防控制所虫媒室, 北京 102206
  • 收稿日期:2021-05-17 出版日期:2021-08-20 发布日期:2021-08-20
  • 通讯作者: 王环宇,E-mail:wanghy@ivdc.chinacdc.cn
  • 作者简介:何英,女,主管技师,主要从事虫媒病毒相关研究工作,E-mail:hey@ivdc.chinacdc.cn;殷启凯,男,硕士,研究实习员,主要从事虫媒病毒病原学的相关研究,E-mail:yinqk@ivdc.chinacdc.cn
  • 基金资助:
    国家科技重大专项(2018ZX10734-401-001,2018ZX10711001);传染病预防控制国家重点实验室课题(2015SKLID505)

Establishment of a method for determining the virus titer of the Culex flavivirus isolated in China

HE Ying, YIN Qi-kai, FU Shi-hong, WANG Huan-yu   

  1. Department of Arboviruses, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China
  • Received:2021-05-17 Online:2021-08-20 Published:2021-08-20
  • Supported by:
    Supported by the National Science and Technology Major Project of China (No.2018ZX10734-401-001, 2018ZX10711001) and the Development Grant of State Key Laboratory of Infectious Disease Prevention and Control (No. 2015SKLID505)

摘要: 目的 建立我国分离库蚊黄病毒(Culex flavivirus,CxFV)毒株(SDDM06-11)的病毒滴度测定方法。方法 在6孔细胞培养板中培养C6/36细胞,感染库蚊黄病毒,加入甲基纤维素培养4 d,结晶紫进行染色,通过统计空斑数量计算病毒的噬斑形成单位。结果 使用甲基纤维素-结晶紫空斑方法观察,发现CxFV(SDDM06-11)毒株感染C6/36细胞后形成清晰的圆形噬斑,大小如针尖,成功测定CxFV(SDDM06-11株)滴度在106.85pfu/ml。结论 甲基纤维素-结晶紫空斑法可用于测定CxFV滴度。

关键词: 库蚊黄病毒, 病毒滴度, 噬斑试验

Abstract: Objective To establish a method for determining the virus titer of the Culex flavivirus (CxFV) strain (SDDM06-11) isolated in China. Methods C6/36 cells were cultured in a 6-well cell culture plate and were then inoculated with CxFV. Methylcellulose was added to the cells and cultured for 4 days. Crystal violet was used for staining, and the number of plaques was determined to calculate the plaque forming unit of the virus. Results Observation of plaques using methylcellulose and crystal violet showed that CxFV (SDDM06-11) strain formed clear round plaques with the size of a needle tip after infecting C6/36 cells, and the virus titer of SDDM06-11 strain was successfully determined as 106.85 pfu/ml. Conclusion The methylcellulose-crystal violet plaque forming assay can be used to determine the titer of CxFV.

Key words: Culex flavivirus, Virus titer, Plaque assay

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