抗环瓜氨酸肽抗体定量检测ELISA试剂盒的制备及其诊断类风湿性关节炎的意义

目的制备抗环瓜氨酸肽(CCP)抗体定量检测ELISA试剂盒,并探讨其诊断类风湿性关节炎(RA)的意义。方法应用作者合成的CCP制备抗CCP抗体定量检测试剂盒,并进行鉴定。应用该试剂盒检测178份RA患者和712份非RA患者及健康人血清,评价其在RA诊断中的价值。结果所制备的定量检测试剂盒cutoff值为25RU/ml;线性良好,R2值大于0.99;试验内变异系数为5.15%~8.25%,试验间变异系数为6.82%~11.23%;平均回收率为99.7%。该试剂盒在4℃保存6个月,稳定性良好。与国外同类试剂盒同时测定80份样本,阳性符合率为95.7%,阴性符合率为97.1%,总体符合率为96.3%。该试剂盒对RA诊断的敏感性为73.0%,特异性为98.2%。结论所制备的试剂盒各项指标均达到相关要求,能够满足临床检测的需要。

preparation of ELISA Kit for Quantitative Determination of Antibody against Cyclic Citrulline Peptide and Its Significance in Diagnosis of Rheumatoid Arthritis

Objective To prepare an ELISA kit for quantitative determination of antibody against cyclic citrulline peptide (CCP) and study its significance in diagnosis of rheumatoid arthritis (RA).Methods An ELISA kit for quantitative determination of antibody against CCP was prepared by using the synthetic CCP with patent sequence and structure,and identified.Determine serum samples from patients with RA,the patients without RA and healthy person to evaluate the significance of the kit in diagnosis of RA.Results The cutoff value of CCP antibody concentration determined by the prepared kit was 25 RU/ml.The determination result was linearly related to CCP antibody concentration.The R2 value of determination result was more than 0.99,and the inter- and intra-CV values were 5.15%～8.25% and 6.82%～11.23% respectively.The mean recovery rate of the kit was 99.7%.The kit showed good stability after storage at 4℃ for 6 months.The positive,negative and total coincidence rates of the prepared and imported kits were 95.7%,97.1% and 96.3% respectively in determination of 80 samples.The sensitivity and specificity of the prepared kit were 73.0% and 98.2% respectively in diagnosis of RA.Conclusion The prepared kit met the relevant requirements and was suitable for the clinical diagnosis of RA.