自制抗人磷脂酶A2受体抗体ELISA试剂盒的临床应用评价

Clinical application of homemade ELISA kit for anti-human phospholipase A2 receptor antibody

摘要:

旨在自制能够辅助诊断原发性膜性肾病的抗磷脂酶A2受体 (PLA2R) 抗体酶联免疫吸附试验 (ELISA)定量检测试剂盒；通过包埋HEK293细胞表达的重组抗原、患者血清与抗原反应、酶的催化放大效应测定样本中抗PLA2R IgG滴度，通过临床数据分析、与市售试剂盒的比对，评价该试剂盒的临床应用价值及性能。研究发现自制试剂盒与国外试剂盒检测阳性一致率97.2%，阴性一致率100%；检测限不高于2 RU/mL；准确度的测量相对偏差在±15%区间内；试剂盒线性范围在2–500 RU/mL，线性相关系数r值不低于0.990 0；重复性检测的变异系数 (CV) 小于15%；于37 ℃恒温箱放置1周后，2–8 ℃放置1年后，检测性能无明显改变。结果表明自制试剂盒对血清抗PLA2R抗体检测的敏感性、特异性均与德国欧蒙公司 (E150522BD) 试剂盒相近，诊断效能高，可用于原发性膜性肾病辅助诊断。

Abstract:

To develop and evaluate an ELISA kit for Anti-PLA2R IgG. Recombinant M-type phospholipase A2 receptor (PLA2R) protein expressed in HEK293 was taken as coating antigen, HRP-labeled rabbit anti-human IgG was taken as a tracer, to test the anti-PLA2R IgG on the basis of the principle of ELISA. The detection linear range, accuracy, linear correlation, repeatability, and stability were evaluated. In addition, we made a comparison with enrolled anti-PLA2R IgG kit. The detection linear range of the kit is no more than 2 RU/mL. The relative deviation of the kit accuracy is in –15%–15%. There is a linear correlation coefficient of higher than 0.990 0 within 2–500 RU/mL range. The CV of the repeatable test is lower than 15% when the kit was put in 37 ℃ one week, 2–8 ℃ one year, the performance remains. The consistency of testing with comparison in enrolled anti-PLA2R IgG kit is 98.9% (positive: 97.2%, negative: 100%). The Anti-PLA2R IgG ELISA Kit which we developed is nearly identical to the reference standard in specific and sensitive of clinics. It’s successfully used to determine anti-PLA2R titer and help clinical diagnosis.