单抗免疫斑点法和组织印迹法检测百合无症病毒

应用抗百合无症病毒（Lily symptomless virus，LSV）的单克隆抗体，建立了快速检测田间样品的免疫斑点法（Dot—ELISA）和组织印迹法（Tissue blot-ELISA）体系。LSV单抗稀释2，000倍时，Dot-ELISA中病叶粗汁液可被检出的最大稀释度为1：640。Tissue blot—ELISA中样品一次平切后第1次印迹与Dot—ELISA样品1：40稀释的结果相当，前4次印迹均可获得满意的显色效果。常规Tissue blot-ELISA的灵敏度低于Dot—ELISA，但用丙酮处理点过样的硝酸纤维素膜后，二者的灵敏度相当，且Tissue blot—ELISA操作更简便，适合田间大量样品的快速检测。

Detection of LSV in Lily Tissue by Dot-ELISA and Tissue Blot-ELISA with Monoclonal Antibody

Dot-ELISA and tissue blot-ELISA with monoclonal antibody to detect Lily symptomless virus(LSV) were established. 2000-dilution LSV monoclonal antibody could be adopted to detect LSV infecting leaf extract within a dilution limitation of 1:640. LSV could still be detected in the forth print by tissue blot-ELISA. Similar results were shown between the first print by tissue blot-ELISA and 40-dilution of infected leaf extract by dot-ELISA. Comparison tests showed that the commonly used tissue blot-ELISA was less sensitive than dot-ELISA,but soaked cellulose nitrate membrane (NCM) with acetone in tissue blot-ELISA produced the same sensitivity as dot-ELISA. 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.