To explore the difference of the effect of three-dimensional ultrasound combined with tomographic ultrasound imaging (TUI) technology and serum β-human chorionic gonadotropin (β-hCG) and progesterone (P) levels of patients for diagnosing their ectopic pregnancy. Methods: A total of 200 patients with the high suspicion of ectopic pregnancy who admitted to hospital from June 2020 to December 2022 were selected in this study. All the patients had received the transvaginal three-dimensional ultrasound and TUI examinations, and the serum β-hCG and P levels of the patients were detected. The clinical and the postoperative pathology results of the patients were used as the gold standard, the clinical value difference of the transvaginal three-dimensional ultrasound and TUI examinations for diagnosing the ectopic pregnancy of the patients was compared. Results: The pathological results was as the gold standard, the overall coincidence rate of the transvaginal three-dimensional ultrasound combined with TUI examinations for diagnosing the ectopic pregnancy of the patients was 92.50%, and the coincidence rates of them were 94.96% and  81.82%. The detection levels of the serum β-hCG and P of the patients with ectopic pregnancy were significantly lower than those of the patients with intrauterine pregnancy (P<0.05). The sensitivity and the specificity of the transvaginal three-dimensional ultrasound combined with TUI examinations for diagnosing the ectopic pregnancy of the patients were 93.17% and 89.74%, and which of the levels of serum β-hCG and P of the patients for diagnosing their ectopic pregnancy were 81.99% and 71.79%. The results of the transvaginal three-dimensional ultrasound combined with TUI examinations for diagnosing the ectopic pregnancy showed that there were significant differences in the mass diameter, boundary and internal structure, the yolk sac detection rate, and the original heart tube detection rate among the patients with different types of tubal pregnancy (P<0.05). Conclusion: Compared with those of the serum β-hCG and P levels, the transvaginal three-dimensional ultrasound combined with TUI examinations for diagnosing the ectopic pregnancy of the patients has higher sensitivity and specificity, and which can make early relevant judgments for the type of the tubal pregnancy.