To investigate the diagnostic value of transvaginal real-time three-dimensional hysterosalpingo-contrast sonography (RT-3D-HyCoSy) combined with transvaginal two-dimensional ultrasound (TDU) for women with infertility caused by the tubal factors. Methods: 90 infertility women caused by the suspected tubal factors were selected from December 2020 to December 2023 as the research subjects. According to the different examination methods, these women were divided into control group (45 cases with transvaginal TDU) and experimental group (45 cases with transvaginal RT-3D-HyCoSy combined with TDU). The pain degree evaluated by visual analogue scale (VAS) and the occurrence of adverse reactions of the women in the two groups during the two examinations were recorded. The diagnostic value of RT-3D-HyCoSy examination for the infertility of the women was analyzed. Results: The examination time (15.08±2.07 min) and the VAS score (3.01±0.54 points) of the women in the experimental group were significantly lower than those (21.12±2.68 min and 5.16±0.78 points) of the women in the control group. The total incidence of adverse reactions (2.2%) of the women in the experimental group was significantly lower than that (26.7%) of the women in the control group, and the diagnostic accuracy (97.8%) of the women in the experimental group was significantly higher than that (75.6%) of the women in the control group. The misdiagnosis rate (2.2%) and the missed diagnosis rate (0) of the women in the experimental group were significantly lower than those (15.6% and 8.9%) of the women in the control group (all P<0.05). Conclusion: The transvaginal RT-3D-HyCoSy combined with TDU for diagnosing the infertility of the women is superior to that of the transvaginal TDU, and which can also short the examination time, relieve the pain and reduce the incidence rates of adverse reactions of the women during examination.