Breeding of cultivars resistant to anthracnose is one of the most important grape-breeding goals in Korea. Culture filtrates produced by E. ampelina were used to determine varietal susceptibility to anthracnose in grape cultivars as a substitute for pathogen inoculation or field screening in this study. Resistance evaluated by bioassay of grape leaves with culture filtrates and their ethyl acetate extracts was compared with ones from pathogen inoculation and field screening. To evaluate the resistance to anthracnose disease in grape germplasm, European grapes, American grapes, and Vitis hybrids were tested. Bioassay with culture filtrates produced by the pathogen showed that `Black Eye', `Mario', `Niunai', `Rizamat', and `Rosario Bianco' were sensitive; and `Campbell Early', `Niagara', and `Honey Red' were tolerant to anthracnose. In the result of anthracnose resistance evaluation by pathogen inoculation, some cultivars, such as `Black Swan','Rizamat', `Rosario Bianco', and `Kaiji', were susceptible; and others, such as `Campbell Early', `Niagara', `Sheridan', and `Izumo Queen', were found to be resistant to anthracnose. Evaluation in the vineyard showed that `Black Eye', `Mario', `Niunai', `Rizamat', and `Rosario Bianco' were susceptible; `Campbell Early', `Niagara', and `Honey Red' were resistant. The results of bioassay with culture filtrates of the pathogen were consistent with ones from the results by pathogen inoculation and screening in the vineyard.