Objective: To compare the diagnostic value of biopsy under percutaneous transhepatic cholangiography (PTC) versus spiral CT in obstructive jaundice. Methods: The clinical records of 78 patients admitted from 2010 March to 2012 December due to obstructive jaundice were collected. All patients underwent either PTC biopsy or spiral CT examination, and the two diagnostic methods were compared according to the results of the surgical pathology biopsy. Results: Of the 78 patients, 9 cases were identified as benign obstructive jaundice, and 69 cases were diagnosed as malignant obstructive jaundice that included 46 biliary epithelial tumors and 23 non-biliary epithelial tumors. The sensitivity and specificity of PTC biopsy was 85.51% and 66.67%, and those of spiral CT was 78.26% and 77.78%, respectively. The sensitivity of PTC biopsy was higher, but its specificity was lower than that of spiral CT, and both differences were statistically significant (both P<0.05). In the 46 patients with bile duct cancer, 43 cases were positive by PTC biopsy (93.48%) and 37 cases were positive by spiral CT (80.43%); in the 23 patients with non-bile duct cancer, 19 cases were positive by PTC biopsy (82.61%), and 17 cases were positive by spiral CT (73.91%). The sensitivity of PTC biopsy in these patients was higher than that of spiral CT (P<0.05), and further, the sensitivity of PTC biopsy for biliary epithelial tumors was higher than that for non-biliary epithelial tumors (P<0.05), while no such difference was noted in spiral CT for the tumors of the two origins (P>0.05). Conclusion: The sensitivity of PTC biopsy for diagnosis of different types of obstructive jaundice, especially those due to bile duct cancer, is higher than that of spiral CT, but it has many contraindications, so other different methods also can be used according to the clinical needs.