Three viroids: Citrus exocorlis viroid (CEVd), Hop stunt viroid (HSVd), and Potato spindle tuber viroid (PSTVd) can be detected in naturally infected grapevine by biological indexing and molecular methods. In the present study, direct tissue-printing and dot-blot on the membranes, procedures has been applied on a large scale for an initial screening of HSVd, CEVd and PSTVd in Egypt. The results showed that the tissue print assays allowed clear discrimination between healthy and viroid-infected grapevine plants than dot-blot hybridization, the number Of HSVd-infected grapevine plants were 10 plants, PSTVd were 10 plants and the CEVd was detected with high incidence level in grapevine, where 12 out of 100 grapevine trees analyzed were infected. The frequency of HSVd, CEVd and PSTVd naturally infected grapevine trees recorded different percentages as in single (8.33, 8.33 and 11.11%) respectively, double (11.11, 11.11 and 19.44%) for CEVd+HSVd, CEVd+PSTVd and HSVd+PSTVd respectively and mixed infection (19.44%) with different disease symptoms. The pervious results illustrated that, tissue printing hybridization was more reliable than dot-blot hybridization in viroid detection. HSVd was isolated on Cucumis sativus L. cv. Alpha plants which showed specific symptoms severe mosaic, vein clearing, rugosity and yellowing spots. CEVd was isolated on Gynura aurantiaca plants which showed specific symptoms mild mosaic and mottling and Lycopersicon esculantum L. cv. Castle rock reacted with PSTVd producing leaf curl and epinasty.