Preparation And Evaluation Of Self-made Quality Control Products Against APF,AKA And Anti-CCP Antibodies

Background Rheumatoid arthritis(RA)is a very common chronic inflammatory disease characterized by progressive symmetrical joint swelling,joint tenderness,and synovial joint destruction,which can lead to severe disability and premature death.Early diagnosis and active treatment of RA play a very important role in alleviating joint damage and improving the quality of life of RA patients.Antiperinuclear factor antibody(APF),Antikeratin antibody(AKA),anti-Cyclic citrullinated pepide(CCP)antibody these three autoantibodies have high specificity,which can be used to predict the prognosis of RA in early diagnosis and widely used in clinic.However,as a highly heterogeneous protein,antibodies has always been a great challenge in the standardization of detection.The determination of autoantibodies is mainly performed by manual manipulation and human eyes,which increases the uncontrollability of results.In the absence of standards,laboratory testing requires more quality control to ensure the control of the testing process and the quality of the results.However,there are few indoor quality control products on the market and they are expensive,so a kind of quality control products with affordable price and good performance is urgently needed to meet the requirements of laboratory testing.Objective To investigate the feasibility of self-made quality control products with APF,AKA and anti-CCP antibody.It provides reference for the preparation of other autoantibodies quality control products.At the same time for the laboratory detection of autoantibodies to find easy access to available quality control products.Methods Collect the serum of RA patients whose three autoantibodies are positive for APF,AKA and anti-CCP antibody and the negative serum of healthy people on physical examination,and mixed in proportion to make the required concentration of mixed serum.The mixed serum was inactivated by heating in 56 ? for 30 minutes,then centrifuged and filtered,and stored in a refrigerator below-20?.20 parts of the mixed serum were extracted,and one part were tested 20 times and 20 parts of each were tested once to detect the difference between bottles of the mixed serum and evaluate the uniformity of the mixed serum.Stability at 37? was determined by testing every 24 hours under water bath conditions.Under-20? freezing condition,tested once a week to determine the long-term stability of the mixed serum.Results In the test of inter-bottle difference,APF and AKA detected no inter-bottle difference.The external bottles difference CV of anti-CCP antibody was 3.2%,far less than the requirement of 20%,showing a good uniformity.Under 37? water bath for 8consecutive days,APF and AKA results were within the acceptable range of the next titer,and the maximum bias of anti-CCP antibody was-15.8%,less than the ±20%bias requirement.In the long-term stability experiment of-20 ?,after 9 months of detection,the detection results of APF and AKA were all within the acceptable range of the next titer,the monthly CV fluctuation of anti-CCP antibody was all less than 20%,and the total CV was 10.6%,less than the requirement of 20%.All negative quality control products tested negative.Conclusion The self-made mixed serum quality control products of APF,AKA and anti-CCP antibody have good performance,which can meet the requirements of laboratory quality control.And the method is simple,maneuverable and applicable to all levels of hospitals and laboratories.