Establishment And Evaluation Of Magnetic Particles Chemiluminescence Quantitative Determination Of Anti-cyclic Citrullinated Peptide Antibody

Objectives: A magnetic particle chemiluminescence immunoassay for the quantitative determination of anti-cyclic citrullinated peptide antibody(Anti-cyclic citrullinated peptide antibody,CCP)was established,and its performance evaluation and methodological comparison were carried out.Methods: 1.The cyclic citrullinated peptide antigen(Cyclic citrullinated peptide,CCP),was labeled with biotin and the concentration of biotinylated antigen was determined by BCA method.2.To explore the optimum concentration of biotinylated antigen,streptavidin magnetic beads and dilution ratio of HRP labeled antibody in the detection system.Incubation time.3.The sensitivity,specificity and precision of the method were evaluated and compared.Results: 1.The concentration of biotinylated antigen determined by1.BCA method was 0.3mg/ml,which indicated that the antigen labeling was successful and the follow-up experiment could be carried out.2.The results showed that the optimum concentration of streptavidin magnetic beads was0.75?g/ml;,the dilution ratio of streptavidin magnetic beads was 0.225?g/ml;HRP,and the reaction time was 15min+20min.3.Through the evaluation of performance parameters,the detection limit(MDL)and quantitative detection limit(QDL)were 11.21RU/ml and 36.99RU/ml,respectively.There was no obvious cross reaction to high concentration of anti-U1-nRNP antibody,anti-SS-A antibody,anti-dsDNA antibody,anti-nucleosomeantibody(NUC)and anti-RO-52 antibody.The repeatability was2.7%-8.5%.4.Compared with Roche,the results showed that the correlation was good,y=0.83x+38.412,R2 = 0.901(P < 0.001).Conclusion: 1.After a series of adjustments,such as antigen labeling,antigen coating,antigen magnetic bead concentration screening,optimization and so on,this method has the ability to determine anti-CCP antibody in clinical samples.2.Through systematic performance evaluation and methodological comparison,this method has the ability to quantitatively detect the concentration of anti-CCP antibody in clinical samples,and can be used for clinical detection and research.