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林业科学 ›› 1999, Vol. 35 ›› Issue (1): 60-65.

• 论文及研究报告 • 上一篇    下一篇

马尾松毛虫质型多角体病毒单克隆抗体的制备和应用

朱光旦1,林军1,沈中建1,钟江1,陈昌洁2   

  1. 1. 复旦大学生命科学学院病毒研究室 上海 200433
    2. 中国林业科学院森林保护研究所 北京 100091
  • 收稿日期:1996-07-23 出版日期:1999-01-25 发布日期:1999-01-25

PREPARATION AND APPLICATION OF MONOCLONAL ANTIBODIES AGAINST DENDROLIMUS PUNCTATUS CYTOPLASMIC POLYHEDROSIS VIRUS

Changdan Zhu1,Jun Lin1,Zhongjian Shen1,Jiang Zhong1,Changjie Chen2   

  1. 1. Virology Research Unit, School of Life Sciences, Fudan University Shanghai 200433
    2. Chinese Academy of Forestry Beijing 100091
  • Received:1996-07-23 Online:1999-01-25 Published:1999-01-25

摘要:

纯化的马尾松毛虫质型多角体病毒(DpCPV)云南文山株的病毒粒子经SDS-PAGE分析,其结构蛋白有120kd、116kd、110kd、66kd和33kd5个组分,而它的多角体蛋白含30kd和28kd两个主要组分。以纯化的DpCPV粒子为抗原制备了2D5、2D10、3D4和6B3共4种单克隆抗体,并测定了它们的亚类。制得的单克隆抗体用于DpCPV的ELISA检测。对DpCPV在棉铃虫卵巢细胞系SFE-HA-8212和SFE-HA-831中增殖动态的检测结果表明, DpCPV感染培养细胞具有释放病毒量小和呈现持续感染等特点。用Western印迹法在SFE-HA-831细胞感染DpCPV后第18h检测到病毒抗原的合成。运用所建立的免疫学检测方法,对几批采自DpCPV防治林区的幼虫样品进行分析,结果表明,该方法适用于对DpCPV的防治效果及其自然流行进行长时期、大规模的监测。

关键词: 马尾松毛虫质型多角体病毒, 单克隆抗体, 生物防治

Abstract:

Five structural polypeptides, with molecular weights of 120 kd, 116 kd, 110 kd, 66 kd and 33 kd, respectively, were found in SDS-PAGE profile of purified virus particles of Dendrolimus punctatus cytoplasmic polyhedrosis virus (DpCPV), Wenshan (Yunnan Province) strain. DpCPV polyhedron contains two major polypeptides of m.w. 30 kd and 28 kd. Four monoclonal antibodies, as 2D5, 2D10, 3D4 and 6B3, were prepared against purified DpCPV particles, and their subclasses were determined. These monoclonal antibodies were used in DpCPV detection by ELISA. Results obtained in vitro DpCPV replication in SFE-HA-8212 and SFE-HA-831 cells (ovarian cell lines from Helicoverpa armigera) showed that these cells were persistently infected, however, the release of virus particles into the culture medium was seldom demonstrated. In Western blotting, traces of viral antigens were detected in SFE-HA-831 cells 18 hours after DpCPV infection. Batches of Dendrolimus larvae collected from pine forests in Guangdong and Yunnan provinces were examined by the immunological detection technique described here. Results confirmed its efficacy in long-term monitoring of DpCPV in Dendrolimus control and its epidemics in nature.

Key words: Dendrolimus punctatus cytoplasmic polyhedrosis virus, Monoclonal antibody, Biological control