Development of Dot-immuno-bloting technique for anti-CCP antibody determination and its preliminary application

Objective To establish Dot-immuno-bloting(DIB) technique for anti-Cyclic citrullinated peptide(anti-CCP) antibody,and study its value in diagnosing rheumatoid arthritis(RA).Methods By optimizing experimental conditions of DIB for antiCCP antibody determination,serum levels of anti-CCP in patients with RA,other autoimmunity and medicine diseases as well as normal control was investigated,and was also determined by ELISA(Based on our established method and EUROIMMUN assay).Results The diagnostic sensitivity of anti-CCP antibody for RA was 52%,and specificity was 94% by this DIB technique.In all patients with RA(n=48),25 of them were positive with anti-CCP antibody,they were significantly increased than other diseases and normal control(P0.01).Parallel investigation showed that serum levels of anti-CCP antibody detected by DIB was highly significant association(χ2=131.712) and consistency(Kappa=0.771) with of ELISA method established based on RSA-CCP immobilization(P0.001),and was also significant association(χ2=24.147) and consistency(Kappa=0.714) with of EUROIMMUN assay(P0.001).Conclusion DIB technique used for anti-CCP antibody determination was successfully established,results of this assay was stable,specific and reliable.So,this DIB technique is worth of popularizing and further investigation.