A novel strategy for cloning and expression of Auto-antigen trimer of M2 Anti-mitochondrial antibody / 国际检验医学杂志

ABSTRACT

Objective To modify recombinant plasmid construction of BPO trimer,and establish the detec-tion of M2 type anti-mitochondrial antibody (AMA-M2) using ELISA.To modify the recombinant expression vector of the target gene triplet of AMA-M2,and to establish and evaluate an enzyme linked immunosorbent assay (ELISA) for the detection of AMA-M2.Methods The expression vector pGEX 4T 1 BPO was con-structed through the gene amplification of triplet BPO by polymerase chain reaction (PCR),and expressed in E.coli BL21,and the recombinant protein was induced by isopropyl Thioglucoside (IPTG).The target protein was purified by affinity chromatography,and ELISA was established to detect AMA-M2.Results The recom-binant protein with a purity greater than 95% was obtained.326 cases of clinical samples was examined,and in 43 cases of primary biliary cirrhosis (PBC),41 cases showed AMA-M2 positive,the sensitivity was 95.3%(41/43),the specificity was 98.2%(278/283),and the difference of AMA-M2 detection rate between PBC group and non PBC group was statistically significant.Conclusion The target protein obtained by the im-proved technology can be used for AMA-M2 detection.AMA-M2 has high sensitivity and specificity for the clinical diagnosis of PBC.