Development of an enzyme immune assay for detecting M2 autoantibodies specific for primary biliary cirrhosis.
OBJECTIVE
To develope a new enzyme immune assay (ELISA) for detection of M2 antibody specific for primary biliary cirrhosis (PBC) by using a triple hybrid clone as antigen, which coexpresses the three immunodominant lipoyl domains of PDC-E2, BCOADC-E2 and OGDC-E2 from human sources.


METHODS
After expressing autoantigens of PBC in prokaryote by constructing recombinant expressive plasmid successfully, the fusion protein was purified by affinity chromatography. The sera of 17 PBC patients were examined. As controls, the sera of 167 non-PBC patients and the sera of 1225 normal controls aged under 28 were examined.


RESULTS
None of the sera from the non-PBC patients or the normal controls was positive for anti-M2 shown by the new ELISA. However, the positivity rate for anti-M2 in the PBC patients was 100% (17/17), as shown by the new ELISA.


CONCLUSIONS
The detection system with a good sensitivity and specificity may be used as a powerful method for the diagnosis of PBC.
Jiang
Xiao-Hua
XH
Department of Laboratory Medicine, 85th Military Hospital, Shanghai 200052, China. jhlulu@citiz.net
Fang
Xiao-Yun
XY
Zhong
Ren-Qian
RQ
Wang
Xiu-Ping
XP
Hu
Yin
Y
Kong
Xian-Tao
XT
D000328:Adult; D001323:Autoantibodies / Q000032:analysis* / Q000097:blood / Q000276:immunology; D001324:Autoantigens / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D000939:Epitopes; D015870:Gene Expression / Q000276:immunology; D006801:Humans; D016056:Immunodominant Epitopes / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D008105:Liver Cirrhosis, Biliary / Q000175:diagnosis* / Q000276:immunology*; D017434:Protein Structure, Tertiary
D001323:Autoantibodies; D001324:Autoantigens; D000939:Epitopes; D016056:Immunodominant Epitopes