Data on the prevalence of strongyloidiasis in Ecuador are patchy. The aim of this study was to document the presence of Strongyloides stercoralis infection in rural communities of different provinces of Ecuador. We tested 1,418 serum samples stored at the biobank of the Central University of Ecuador, Quito, with an ELISA test for Strongyloides. The samples had been collected in eight different provinces of Ecuador. Two hundred ninety-four samples (20.7%) were positive, and Jipijapa, Manabí Province, was the site with the largest proportion of positive samples (66.7%). Further surveys aimed at estimating the prevalence of the infection should be carried out in areas where the infection seems highly prevalent, and ad hoc control measures should be adopted.