Objective: The aim of this study was to suggest a new sonographic sign, which could be helpful in the determination of chorionicity in twin pregnancies, during the second and third trimesters. Materials and Methods: This was a retrospective study evaluating the sonographic images of 50 women with twin pregnancy. Out of 50 twin pregnancy case reviewed, only 27 were included. Those patient cases that were included were scanned by a single radiologist. The presence of a hypoechoic cleft, in the thickness of placental bulk, could be detected in all of the dichorionic pregnancy cases. This sonographic appearance was referred to as the “cleft sign.” Results: The “cleft sign” was present in all the dichorionic diamniotic (DC-DA) twin pregnancy cases, but no similar appearance was seen in the monochorionic diamniotic (MC-DA) twins. Considering this sign, the radiologist was able to correctly diagnose the chorionicity of all 24 patients in the second and third trimester. Conclusion: The findings of the present study suggest a possible role for this newly identified “cleft sign.” This sonographic appearance is a hypoechoic cleft in the thickness of placenta, of dichorionic pregnancies, as an indicator of DC twin pregnancy. In this retrospective case analysis, the “cleft sign” was diagnostically significant in the second and third trimesters.