Fusarium wilt is a serious disease of cucurbit crops including cultivated Luffa species (Luffa aegyptiaca, Luffa acutangula) causing considerable amount of reduction in yield and quality. Luffa is starting to be used as rootstocks for major commercial cucurbit crops, but little is known of its resistance against soilborne diseases. Here, 63 Luffa accessions from the World Vegetable Center genebank were evaluated for resistance to an aggressive isolate of Fusarium oxysporum f. FoCu-1 (Fsp-66). According to visual screening based on disease severity rating, 14 accessions exhibited a high level of resistance against Fsp-66. These accessions were further evaluated for resistance against Fsp-66 and two more isolates FoCu-1 (isolated from infected cucumber plants) and FoM-6 (isolated from infected bitter gourd plants). Of the 14 accessions, 11 were confirmed resistant against isolate Fsp-66. In addition, 13 accessions showed high resistance against isolates FoCu-1 and FoM-6. This is the first report of Fusarium wilt resistance in Luffa and these sources will be valuable for the development of Luffa rootstocks/cultivars resistant to soil-borne pathogen to manage this serious disease.