摘要
用香石竹斑驳病毒(CarMV)免疫的BALB/c鼠脾细胞与Sp2/0鼠骨髓瘤细胞融合,经筛选克隆,获得5株能稳定传代且分泌抗CarMV单克隆抗体(MAb)的杂交瘤细胞,并分别制备它们的单克隆抗体腹水。5株单克隆抗体腹水间接ELISA效价达10-6,其中3G1、1B9、2A9和2F8的抗体类型及亚类均为IgG1,而2F2为IgG3。W estern-b lot分析表明,5株单克隆抗体均与CarMV 38 kD的外壳蛋白亚基有特异性反应。利用2A9单抗建立的抗原包被的间接ELISA(ACP-ELISA)检测CarMV方法,病叶1∶800倍稀释、提纯CarMV病毒浓度为1 ng/mL(绝对检测量为0.1 ng)时仍能检测到病毒。利用ACP-ELISA对田间香石竹样品的检测表明,CarMV在香石竹上发病很普遍。
Five hybridoma cell lines secreting monoclonal antibodies （MAbs） against Carnation monle virus （CarMV） were produced by fusing mouse myeloma cells （Sp2/0） with spleen cells from BALB/c mice immunized by the CarMV particles. The five MAbs could specifically react with CarMV. The titres of ascitic fluids of the five MAbs are 10-6 in indirect antigen-coated plate （ACP）- ELISA. Isotypes and subclasses of 3G1,1B9,2A9 and 2R3 belong to IgG1 while that of 2F2 belong to IgG3. Western-blot analysis showed that all the five MAbs can react specifically with the 38 kD coat protein subunit of CarMV. The subclass 2A9 was used in ACP-ELISA for detection of CarMV. ACP-ELISA could successfully detect 0.1 ng purified CarMV or virus in plant sap diluted 800 fold. The presence of CarMV in field Dianthus caryophyllus tissues was investigated with ACP-ELISA and the detection results showed that CarMV is common in field samples of Dianthus caryophyllus.
出处
《植物病理学报》
CAS
CSCD
北大核心
2005年第4期322-326,共5页
Acta Phytopathologica Sinica
基金
浙江省科技厅(成果转化)项目(G20030724)
国家高技术研究发展计划(2003AA2Z2091)
国家自然科学基金资助项目(30370305)
