抗环瓜氨酸肽抗体胶体金定量检测方法的建立

目的  建立一种检测人血清中抗环瓜氨酸肽（CCP）抗体含量的胶体金定量检测方法。方法  采用间接胶体金技术，对胶体金最佳标记抗体pH、最低标记抗体量、最佳T线包被浓度、金标定量仪最佳读数时间、批内批间精密性、回收率、临床标本检测等研究，建立抗CCP抗体胶体金定量检测方法并评价其性能。结果  实验确定胶体金标记最佳pH及最低标记抗体量为1 ml胶体金溶液中加入0.2 mol/ml 碳酸钾K2CO3 3μl、鼠抗人IgG 8.8μg；T线包被浓度选择CCP-BSA偶联抗体1.5 mg/ml；使用金标定量仪时，最佳读数时间为14 min。性能评价结果显示建立的检测方法线性范围为50～500 RU/ml，R2=0.993；精密性质控品L1批内CV= 2.6%，批间CV=4.52%；精密性质控品L2批内CV=2.5%，批间CV=4.28%；高、中、低值血清的回收率为100.96%、97.17%、91.46%；与国外试剂盒平行检测40份抗CCP抗体临床血清，结果显示两者相关性较好，回归方程：Y=1.004X+7.968，R2=0.955。结论  初步建立一种检测人血清中抗CCP抗体含量的胶体金定量检测方法，该方法具有较高的准确性和精密性。

Establishment of colloidal gold quantitative detection method of anti-CCP antibody

Objective To establish a colloidal gold quantitative method for detection of anti-CCP antibody levels in human serum. Methods Indirect colloidal gold technique was used to detect colloidal gold labeled antibody best pH, minimum amount of labeled antibody, the best package concentration of T line, optimal reading time of Colloidal gold quantitative instrument, precision, recovery and clinical samples. Anti-CCP antibody colloidal gold quantitative detection method was established and its performance was evaluated. Results The colloidal gold marked optimal pH and minimum amount of labeled antibody was 1 ml colloidal gold solution with 0.2 mol/ml K2CO3 3 μl, rat IgG 8.8 μg; T lines has been selected CCP-BSA coupling antibody 1.5 mg/ml. The best reading time of colloidal gold quantitative meter was 14 min. Linear ranged 50~500 RU/ml, R2 = 0.993. Conclusions A quantitative colloidal gold kit for detection of anti-CCP with high performance is established.