单抗免疫斑点法和组织印迹法检测侵染蝴蝶兰的建兰花叶病毒

Detection of CymMV in Orchids Tissue by Dot-ELISA and Tissue Blot-ELISA with Monoclonal Antibody

摘要:

应用抗建兰花叶病毒(CymMV)的单克隆抗体, 建立了快速检测蝴蝶兰病样的免疫斑点法(Dot-ELISA)和组织印迹法(Tissue blot-ELISA)。CymMV单抗稀释8000倍时, Dot-ELISA可检出病毒粗汁液的最大稀释度为1:10240; Tissue blot-ELISA中样品1次平切后1~5次印迹与Dot-ELISA样品1:80稀释结果相当, 6~8次印迹与Dot-ELISA 1:320稀释结果相当, 前8次印迹均可以得到满意的检测效果。Tissue blot-ELISA的灵敏度略低

Abstract:

Dot-ELISA and tissue blot-ELISA with monoclonal antibody to detect Cymbidium mosaic virus (CymMV) were established. 8000-dilution CymMV monoclonal antibody could be adopted to detect CymMV infecting leaf extract within a dilution limitation of 1:10240. CymMV could still be detected in the eighth print by tissue blot-ELISA . Similar results were shown between the fifth print by tissue blot-ELISA and 80-dilution of infected leaf extract by dot-ELISA. Comparison tests showed that the commonly used tissue blot-ELISA was less sensitive than dot-ELISA. It could illustrate that CymMV monoclonal antibody was more specific and sensitive than the polyclonal antibodies. The improved tissue blot-ELISA is more simple than dot-ELISA, and is suitable for fast detection with a large number of samples in the field.