Objective: Echogenic intracardiacs focus (EIF), a microcalcification of the papillary muscles in either or both ventricles of the fetal heart, is one of the more common ultrasonographic soft markers of trisomy 21. This study aimed to determine the relationship of EIF with trisomies in the presence of various ultrasonographic findings. Materials and Methods: A retrospective study of second-trimester obstetric sonograms (18-24 weeks) was conducted at a tertiary care center over a two-year period. The patients were divided into three groups: isolated EIF (group 1), EIF with at least one soft marker (group 2), and EIF with structural anomalies (group 3). All the groups were divided into subgroups on the basis of a screening test (maternal age ≥ 35, positive combined-triple-quadruple tests). The incidence of chromosome anomalies was evaluated. Results: The authors examined 8,300 patients during the study period and found 170 fetuses with isolated EIF (group 1), 26 fetuses with EIF and soft markers (group 2), and 37 fetuses with EIF and structural anomalies (group 3). Thirteen (8%) patients underwent fetal karyotyping in group 1, 10 (38%) in group 2, and 22 (60%) in group 3. The rate of the invasive test was higher in fetuses exhibiting EIF accompanied by at least one of the soft markers. No trisomy was detected in group 1 or 2. Conclusion: The risk of aneuploidy did not increase in either isolated EIF or in cases with other soft markers but only in cases with structural anomalies.