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

作　　者：黄莉敏[1] 张大准 王艳[1] 张巧 梁伟[1] 张永顶 熊祖应[1] Limin Huang1, Dazhun Zhang2, Yan Wang1, Qiao Zhang2, Wei Liang1, Yongding Zhang2, and Zuying Xiong1(1 Department of Nephrology, Peking University Shenzhen Hospital, Shenzhen 518035, Guangdong, China ;2 Shenzhen Blot Biotech Co. Ltd, Shenzhen 518054, Guangdong, Chin)

机构地区：[1]北京大学深圳医院肾内科,广东深圳518035 [2]深圳市伯劳特生物制品有限公司,广东深圳518054

出　　处：《生物工程学报》2018年第5期743-751,共9页Chinese Journal of Biotechnology

基　　金：国家科技支撑计划(No.2013BAI07B01);广东省科技计划项目(No.2014A020212049)资助~~

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。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.

关 键 词：磷脂酶A2受体 抗体 酶联免疫吸附试验 原发性膜性肾病 血清 

分 类 号：R446.6[医药卫生—诊断学] R692[医药卫生—临床医学]