摘要
植物病毒的酶联免疫吸附分析检测技术（ELISA）的反应可以在硝酸纤维膜（NCM）上进行，称为dot－ELISA。为进一步简化实验步骤，改膜上点样为病组织直接印迹，以后则参照间接ELISA程序进行。成功地进行了香石竹斑驳病毒（CarMV）的快速检测，约比常规dot－ELISA缩短时间3h，灵敏度与聚乙烯板上的间接ELISA相似。讨论了使用辣根过氧化物酶标记抗体存在非特异性反应的可能性及克服办法。
Enzyme-linked immunosorbent assay(ELISA) used for detecting plant virus-es has been successfully performed on cellulose nitrate membrane (NCM ),the so called dot-ELISA,which needs a pretreatment of unknown antigen. In order to make dot-ELISA more simple and practical,we adopted direct blotting on NCM by infected tissues instead of dot sampling by pretreated leaf extract,and then performed experiment similarly to dot-ELISA.By this technique,carnation mottle virus (CarMV) was readily detected from leaves,stems,and incompletely opened flowers of carnations. Comparison tests show that direct tissue blot-ting immunoassay is same sensitive as the commonly used ELISA and dot-ELISA for detec-tion of CarMV in extracts or partially purified preparation from infected carnations,and can save 3 h needed in dot-ELISA for pretreatment of extracts. The possibility of nonspecific re-actions from application of HRP-IgG as a conjugated antibody in tissue blotting and the pro-cedure to delete nonspecific reactions are also discussed in the paper.
出处
《上海农业学报》
CSCD
1999年第2期84-86,共3页
Acta Agriculturae Shanghai
基金
上海市青年科技启明星计划资助!94QG14002
