Strongyloidiasis is a soil-transmitted helminthiasis with a high global prevalence. Objectives: We aimed to evaluate the prevalence of Strongyloides stercoralis infection and assess strongyloidiasis serology as a screening technique in the Peruvian Amazon. Material and Methods: We performed a cross-sectional study of strongyloidiasis in 300 pregnant women in Iquitos (Peru) from 1 May 2019 to 15 June 2019. Women were tested using serology (Strongyloides IgG IVD-ELISA kit) as an index test and the modified Baermann technique and/or charcoal fecal culture as the parasitological reference standard. Results: The reference tests showed S. stercoralis in the stool of 30 women (prevalence: 10%; 95% confidence interval [CI] 7.1% to 13.9%), while 101 women tested positive on the blood test (prevalence: 33.7%; 95% CI 28.6% to 39.4%). Fourteen of the 15 women (93.3%) with positive results according to the modified Baermann technique, and 14 of the 23 women (56.5%) with positive charcoal cultures also had positive serological results. Serology showed a sensitivity of 63.3% and a negative predictive value of 94.4%. Conclusion: In Iquitos, pregnant women have a high prevalence of S stercoralis. S. stercoralis ELISA could be an excellent tool for population-based screening, as it has a high negative predictive value that can help to rule out the presence of active infection.