Background: Heterotopic pregnancy is rare in natural pregnancy. In recent years, with the development of assisted reproductive technology, the incidence of heterotopic pregnancy is increasing. Heterotopic pregnancy not only affects the development of intrauterine embryo, but also threatens the life of patients. There is no unified standard for the treatment of heterotopic pregnancy until now. So we research the clinical features and the effect of different intervention time on pregnancy outcome in patients with heterotopic pregnancy (HP) after assisted reproductive technology (ART).Methods: A retrospective analysis was conducted on the patients who were diagnosed as HP in Shanghai first maternal and infant hospital after conception through ART from January 2014 to June 2019. We analyzed the clinical characteristics, therapeutic method and intervention time to explore their impact on the outcome of HP.Results:47 patients with HP after ART were treated in our hospital and have complete clinical data. Among them, 18 cases treated with conservative therapy and 29 cases treated with operation. 29 cases delivered (including 3 cases of pre-term birth and 26 cases of full-term birth) and 18 cases aborted. In HP patients, β Human chorionic gonadotropin (β-HCG, 0.617) and extrauterine mass size (0.242) are not much guiding significance for the treatment. Transvaginal ultrasonography (TVS) examination on admission found that different conditions of ectopic (anechoic area, adnexal mass, heart beat, yolk sac) was independent risk factors of different treatment methods (P=0.005). And the intervention time before six weeks can get better pregnancy outcome (p=0.034).Conclusions:Patients with ART should be examined by TVS as early as possible. If abnormal TVS (anechoic area, adnexal mass, heart beat, yolk sac) results are found, patients should be operated by laparoscopy. And the best intervention time of laparoscopy is before 6 weeks.