In this case-control study, we investigated the seroprevalence and molecular evidence of Chlamydia trachomatis and Waddlia chondrophila in ectopic pregnancies (EP) and uneventful control (C) pregnancies in 343 women from Vietnam. Whereas presence of Chlamydia trachomatis IgG was strongly associated with EP (adjusted Odds Ratio [aOR] 5.41; 95% Confidence Intervals [95%CI] 2.58-11.32), its DNA remained undetected in all tubal lesions. We confirmed an independent association between antibodies against Waddlia and previous miscarriage (aOR 1.87, 95%CI 1.02-3.42). Further investigations are needed to understand the clinical significance of Waddlia high seroprevalence (25.9% in C) in this urban population.