抗环瓜氨酸肽抗体胶体金定量检测方法的建立

发布时间：2018-04-30 23:33

本文选题：抗CCP抗体 + 胶体金　；参考：《中国现代医学杂志》2016年12期

【摘要】：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p H、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p H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K_2CO_33μl、鼠抗人Ig G 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14min。性能评价结果显示建立的检测方法线性范围为50~500 RU/ml,R~2=0.993;精密性质控品L_1批内CV=2.6%,批间CV=4.52%;精密性质控品L_2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,R~2=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。
[Abstract]:Objective to establish a colloidal gold quantitative method for the determination of anti-cyclic citrullinated peptide (CCPP) antibody in human serum. Methods indirect colloidal gold technique was used to study the best labeled antibody pH, the lowest labeled antibody volume, the best T line coating concentration, the best reading time of the gold calibrator, the intra batch precision, the recovery rate, the clinical specimen detection and so on. A method for quantitative determination of colloidal gold with anti-CCP antibody was established and its performance was evaluated. Results the results showed that the best pH labeled by colloidal gold and the lowest labeled antibody were 0.2 mol/ml potassium carbonate K_2CO_33 渭 l in 1 ml colloidal gold solution, and the concentration of CCP-BSA conjugated antibody was selected as the concentration of mouse anti-human IgG 8.8 渭 g / T line coating, and the best reading time was 14 minutes when the gold calibrator was used. The results of the performance evaluation show that the linear range of the established method is 50 / 500 RUP / ml / RJ / 0.993; the precision property controller L1 has a CVN 2.6 in batches and 4.52in interbatch CV52.The precision property controller L2 has a CV2.5 in batches and 4.28 in inter-batch CV2; high, medium, high, medium, and moderate. The recovery rate of low value serum was 100.96 and 97.179.91.46.The results showed that there was a good correlation between the two methods and the regression equation: 1. 004X 7. 96X 7.960.9955.The results showed that the recovery rate of low value serum was 100. 96% and 97. 17% respectively, and 40 clinical sera of anti CCP antibody were detected in parallel with the foreign kits. The regression equation was 1. 004X. Conclusion A colloidal gold quantitative method for the determination of anti CCP antibody in human serum was established. The method has high accuracy and precision.
【作者单位】：郑州大学生命科学学院;郑州职业技术学院;新乡医学院三全学院;河南省生物工程技术研究中心;
【分类号】：R593.22;R446.6

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