Objective To analyze the levels and ratios of carbohydrate antigen 125 (CA125), β-human chorionic gonadotropin (β-hCG), vascular endothelial growth factor (VEGF), creatine kinase (CK), and progesterone (P) of both celiac blood and venous blood of patients with tubal pregnancy, explore the correlation of biomarkers and tubal pregnancy types, and identify biomarkers for the early diagnosis of ruptured tubal pregnancy. Methods The levels of CA125, β-hCG, VEGF, CK, and P of both celiac blood and venous blood of 322 patients with tubal pregnancy were detected. Ratios (Rv/p) of biomarkers of celiac blood and venous blood were calculated. The receiver operating characteristic (ROC) curve was drawn. Best diagnostic thresholds for ruptured tubal pregnancy were determined and their sensitivity and specificity for the diagnosis of ruptured tubal pregnancy were evaluated. Results ①The areas under ROC curve (AUCROC) of CA125v, β-hCGv, Pv, CKv, and VEGFv for the diagnosis of ruptured tubal pregnancy were 0.528, 0.674, 0.570, 0.643, and 0.542, respectively. ②AUCROC of CA125p, β-hCGp, Pp, CKp, and VEGFp for the diagnosis of ruptured tubal pregnancy were 0.896, 0.595, 0.656, 0.586, and 0.682, respectively. ③AUCROC of Rv/p-CA125, Rv/p-(β-hCG), Rv/p-P, Rv/p-CK, and Rv/p-VEGF for the diagnosis of ruptured tubal pregnancy were 0.883, 0.757, 0.607, 0.628, and 0.598, respectively. ④When CA125p<400.15 U/mL, the sensitivity and specificity of diagnosing ruptured tubal pregnancy were 73.68% and 92.83%. ⑤When Rv/p-CA125<15.20, the sensitivity and specificity of diagnosing ruptured tubal pregnancy were 73.68% and 89.81%. Conclusion When CA125p<400.15 U/mL or Rv/p-CA125<15.20, ruptured tubal pregnancy can be diagnosed and the specificity of CA125p<400.15 U/mL is higher.