To investigate the clinical value of the tubal patency through the application of transvaginal real-time three dimensional Hysterosalpingo-contrast sonography (TVS 3D HyCoSy) with SonoVue. In total of 273 tubes of 139 infertility patients was examined via TVS 3D and 2D HyCoSy with SonoVue from November 2011 to May 2014 in Haikou People's Hospital and Shenzhen Bao'an Maternal and Child Health Hospital. Relative data were retrospectively analysed. Under 2D HyCoSy, there were 125 tubal patency, 69 passable, 79 blocked and 24 twisted. Meanwhile, under 3D HyCoSy, there were 122 tubal patency, 73 passable, 78 blocked and 61 twisted. The difference between the evaluation of tubal patency under two modes was not statistically significant (P > 0.05). However, there was significant statistical disparity between the outcomes of the fallopian tube lines under the two modes (P < 0.05), which was assessed better with 3D HyCoSy rather than 2D HyCoSy. Supporting information can be found in the online version of this abstract Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.