Abstract: Objective To evaluate the diagnostic value of anti-mitochondrial antibodies-M2 subtype,  anti-gp210,  anti-Sp100  by  detecting  in  patients  with  primary  biliary cirrhosis (PBC). Methods Serum samples were detected for AMA-M2, anti-gp210 and anti-Sp100 by immunoblotting assay in 2000 patients with cryptogenic hepatitis, and other 15 healthy individuals were taken as control group. Patients were diagnosed as PBC (129 cases, 6.45%), AIH (205 cases, 10.25%), AIH-PBC overlap syndrome (15 cases, 0.75%), fatty liver (514 cases, 25.70%), drug-induced hepatitis (379 cases, 18.96%), and other unexplained hepatitis (758 cases, 37.90%).  Results The positive rates of these three antibodies were 97.67% (126/129), 41.86% (54/129), 27.91% (36/129), respectively. The simultaneous occurrence rate of anti-gp210 and anti-Sp100 was 9.30% (12/129).There were 12 cases with positive anti-gp210 and anti-Sp100 antibody, 3 of whom were diagnosed as PBC through liver puncture. In PBC group, patients with AMA-M2 antibody titer high than 1︰800 accounted for 62.80% (81/129), while the percents of patients with titers ranged from 1︰25 to 1︰100 and from 1︰100 to 1︰200 were 13.95% (18/129) and 30% (12/129), respectively. Conclusions AMA-M2 antibody is still the main marker for PBC diagnosis. The combined detection of AMA-M2 antibody, anti-gp210, anti-Sp100 in PBC have a higher value and play a good supporting role in screening other autoimmune liver diseases.