马尾松毛虫质型多角体病毒单克隆抗体的制备和应用  

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

作　　者：朱光旦[1] 林军 沈中建[1] 钟江[1] 陈昌洁[2] 

机构地区：[1]复旦大学生命科学学院病毒研究室 [2]中国林业科学院森林保护研究所

出　　处：《林业科学》1999年第1期60-65,共6页Scientia Silvae Sinicae

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。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.

关 键 词：马尾松毛虫 质型多角体病毒 单克隆抗体 

分 类 号：S476.13[农业科学—农业昆虫与害虫防治] S763.85[农业科学—植物保护]