Introduction Chagas disease is caused by the protozoan Trypanosoma cruzi. It is endemic in 21 countries in Central and South America. Spain is theonly nonendemic countrywith the highest number of Chagas diseasecases outside the Americas. The only transmission mechanism in Spain is vertical transmission. Materials and methods We reviewed the records of pregnant women from endemic countries who underwent prenatal care at the Hospital Universitario de Guadalajara, from January 1, 2009, to December 31, 2022, to determine the rate of Chagas disease screening and vertical transmission. Results Out of a total of 1,681 pregnant women from endemic countries, prenatal screening was conducted on 316 (18.7%) of them. According to our study, the prevalence of the disease in the population ofpregnant women from endemic countries is 0.95%with a95% confidence interval (ranging from0.32% to2.75%), with three out of the 316 screened women testing positive for the disease. All positive cases were among Bolivian women. Vertical transmission was not observed in any of the cases. However, because of the small sample size, this study cannot conclusively determine the vertical transmission rate in the province of Guadalajara. Conclusions Implementing regulated prenatal screening protocols for Chagas disease at regional or national levels is necessary to increase the rate of prenatal screening. Additionally, increasing awareness of this condition among healthcare professionals and at-risk populations could further improve prenatal screening rates and treatment adherence.