Screening serum markers of primary biliary cirrhosis with high-throughput protein chip encoded by the human genes

Abstract

Objective To screen serum markers in patients with PBC by high-throughput protein chips encoded by the human genome. Methods High-throughput protein chips (contains a total of 38 400protein spots, including 17 718 human genes encoding proteins) were used to screen sera from 21 PBC patients, 20 disease control patients and 10 normal controls. Bioinformatics software was used to analyze information and statistical software was used to analyze the data to confirm the serum markers of PBC. Results The detection rate of protein spots using anti-GST antibody on the chip was 97. 6%, and the signal intensity correlation coefficient of double protein spots was 0. 98. Four serum markers( PDHA1, DBT,DLAT and HK1 )were screened by high-throughput protein chips between PBC group and the control group with a statistically significant. The positive rate of the four markers in the three groups was 66. 67% ( 14/21), 5.00% (1/20) and 0(0/10); 57. 14% ( 12/21 ), 5.00% (1/20) and 0(0/10); 52. 38% ( 11/21 ),0(0/20) and 0(0/10); 52. 38% ( 11/21 ), 0(0/20) and 0(0/10) respectively. All the four markers were different in the three groups with statistically significant (PDHA1 :x2 = 16. 79, P ＜0. 01 ;Fisher exact test,P=0. 000; DBT:x2 =12.86, P＜0. 01;Fisher exact test, P=0. 004; DLAT and HK1:Fisher exact test,P ＜0. 01 or 0. 05). Of those markers, antibodies to PDHA1, DBT and DLAT were the component of AMAM2 which had been used as the marker of PBC. Antibody to HK1 was identified as new marker of PBC,whose sensitivity to PBC was 52. 38% and specificity was 100. 00%. There were no serum marker were screened between the AMA-M2 positive and negative PBC patients. Only antibody to CENPB was identified to be significantly expressed between the ACA positive and negative PBC patients ( Fisher exact test, P =0. 000). Conclusions High-throughput protein chip encoded by the human gene is a technology for quick and comprehensive screening of new markers of PBC. Antibodies to HK1 could be used as new marker for PBC with highly sensitivity and specificity. No serum marker is found between the AMA-M2 positive and negative PBC patients whereas only antibody to CENPB is identified as marker between the ACA positive and negative PBC patients. Key words: Liver cirrhosis, biliary; Protein array analysis; Biological markers; Autoantibodies