肽基精氨酸脱亚胺酶4抗体定性检测试剂盒的研制及初步应用

Preparation and preliminary application of ELISA kit for qualitative determination of antibody against peptidylarginine deiminase 4

目的研制肽基精氨酸脱亚胺酶4(Peptidylarginine deiminase 4,PADI4)抗体定性检测ELISA试剂盒,并初步探讨其对诊断类风湿关节炎(Rheumatoid arthritis,RA)的临床意义及应用价值。方法采用PADI4多肽片段作为包被抗原,制备PADI4抗体定性检测试剂盒,并检测50份RA患者和283份健康人血清,评价该试剂盒的分析性能、精密性和稳定性。使用该试剂盒对181份临床确诊RA患者和353份健康人的血清样本进行初步检测。结果所制备的PADI4抗体定性检测试剂盒总体符合率为85.3%,灵敏度为46.0%,特异性为92.2%;板内变异系数分别为2.47%和4.26%,板间变异系数分别为4.24%和6.20%;在4℃保存6个月,总体符合率、灵敏度、特异性、精密性均无明显变化,稳定性良好。181份RA患者血清中阳性检出率为45.3%,353份健康人血清中阳性检出率为7.7%,差异有统计学意义(P<0.01)。结论所研制的试剂盒可用于类风湿关节炎的临床辅助诊断。

Objective To prepare an ELISA kit for qualitative determination of antibody against peptidylarginine deiminase 4（PADI4） and preliminarily investigate its significance and application in diagnosis of rheumatoid arthritis（RA）.Methods An ELISA kit for was prepared using PADI4 polypeptide as a coating antigen,used for determination of the serum samples from 50 patients with RA and 283 healthy persons,and evaluated for analytical performance,precision and stability.A total of 181 serum samples from patients with RA,as well as 353 from healthy persons,were determined by the developed ELISA kit.Results The general coincidence rate,positive coincidence rate（sensitivity） and negative coincidence rate（specificity） were 85.3%,46.0% and 92.2% respectively.The coefficients of variation of intra-assay of positive and negative samples were 2.47% and 2.46%,while those of inter-assay were 4.24% and 6.20%,respectively.The total coincidence rate,sensitivity,specificity and precision of the kit showed no significant change after storage at 4℃ for 6 months,indicating a high stability.The positive rate of PADI4 in 181 serum samples from patients with RA was 45.3%,while that in 353 samples from healthy persons was 7.7%,which showed no significant difference（P 0.01）.Conclusion The developed kit may be used for assistant diagnosis of RA in clinic.