新型膜性肾病标志物PLA2R抗体的双抗原夹心-时间分辨荧光免疫分析  

Dual-ntigen sandwich-type time-resolved fluoroimmunoassay for phospholipase A2 receptor anti- body, a new marker of membranous nephropathy

作　　者：张艺[1] 胡志刚 王珂[4] 王凉 张秋花 林江[4] 杜明华[5] 顾涛 黄飚[1] Zhang Yi;Hu Zhigang;Wang Ke;Wang Liang;Zhang Qiuhua;Lin Jiang;Du Minghua;Gu Tao;Huang Biao(Key Laboratory of Nuclear Medicine, Ministry of Health, Jiangsu Key Laboratory of Molecular Nuclear Medicine, Jiangsu Institute of Nuclear Medicine, Wuxi 214063, China （Zhang Y, Huang B;Department of Clinical Laboratory, Wuxi People＇s Hospital Affiliated to Nanjing Medical University, Wuxi 214023, China （ Hu ZG;Department of Clinical Laboratory, Jiangyin People＇s Hospital, Wuxi 214400, China （ Wang K, Lin J;Department of Nephrology, Wuxi People＇s Hospital Affiliated to Nanjing Medical University, Wuxi 214023, China （ Wang L, Zhang QH;Department of Nuclear Medicine, Jiangsu Province Hospital of Traditional Chinese Medicine, Nanjing 210029, China （ Du MH;Department of Clinical Laboratory, the First People＇s Hospital of Kunshan, Kunshan 215300, China （ Gu T)

机构地区：[1]江苏省原子医学研究所、卫生部核医学重点实验室、江苏省分子核医学重点实验室,无锡214063 [2]南京医科大学附属无锡人民医院检验科,214023 [3]南京医科大学附属无锡人民医院肾内科,214023 [4]江阴市人民医院检验科,214400 [5]江苏省中医院核医学科,南京210029 [6]昆山市第一人民医院检验科,215300

出　　处：《中华核医学与分子影像杂志》2018年第6期403-407,共5页Chinese Journal of Nuclear Medicine and Molecular Imaging

基　　金：江苏省临床医学科技专项--新型临床诊疗技术攻关项目（BL2014022）;无锡市卫生计生科研项目（MS201642）

摘　　要：目的建立M型磷脂酶A2受体（PLA2R）抗体的时间分辨荧光免疫分析（TRFIA）法，评估该指标在特发性膜性肾病（IMN）中的检测价值。方法以PLA2R抗原包板并对其进行Eu^3＋-链霉亲和素标记，建立PLA2R抗体的双抗原夹心TRFIA法。检测63例IMN患者（男36例、女27例，年龄25～75岁）和90例健康体检者（男30名、女60名，年龄22～53岁）血清中的PLA2R抗体含量。采用Kruskal-Wallis H检验和Mann-Whitney u检验分析数据。结果本方法灵敏度5 μg/L，线性测量范围0～10 mg/L，有效剂量（ED）20、ED50和ED80分别为（0.144±0.012）、（0.707±0.029）和（3.466±0.098） mg/L。批内和批间CV分别为4.7%和5.1%。健康体检者的血清PLA2R抗体水平为0.455（0.320, 0.593） mg/L，而IMN患者的血清PLA2R水平为2.690（0.717, 7.750） mg/L，较前者升高显著（z＝－3.688，P〈0.05）；且血清PLA2R抗体水平在不同年龄组及不同病理分期患者间差异有统计学意义（χ^2值：10.328和9.716，均P〈0.05）。由受试者工作特征（ROC）曲线可知，以血清中PLA2R抗体的质量浓度0.80 mg/L为阈值时，PLA2R抗体检测诊断IMN的灵敏度为73.0%（46/63），特异性为95.6%（86/90）。结论成功建立双抗原夹心测定人血清PLA2R抗体的TRFIA法，该法快速、简便，有助于提高对IMN患者的诊断准确性。ObjectiveTo establish a time-resolved fluoroimmunoassay （TRFIA） method for detecting M-type phospholipase A2 receptor （PLA2R） antibody and to investigate the diagnostic value of serum PLA2R antibody for idiopathic membranous nephropathy （IMN）.MethodsThe microplates coated with recombined PLA2R antigen and Eu^3＋ -streptavidin labeled PLA2R antigen were used to establish a dual-antigen sandwich-type TRFIA for PLA2R antibody detection （anti-PLA2R-TRFIA）. The serum concentrations of PLA2R antibody in 63 IMN patients （36 males, 27 females, age 25-75 years） and 90 healthy volunteers （30 males, 60 females, age 22-53 years） were quantitatively analyzed. Kruskal-Wallis H test and Mann-Whitney u test were used to analyze the data.ResultsThe range of anti-PLA2R-TRFIA was 0-10 mg/L and the sensitivity was 5 μg/L, while ED20, ED50 and ED80 of the standard curve were （0.144±0.012）, （0.707±0.029） and （3.466±0.098） mg/L, respectively. The CV of inter- and intra-assay were 4.7% and 5.1%, respectively. The average concentration of serum PLA2R antibody in healthy volunteers was 0.455（0.320, 0.593） mg/L, but in IMN patients it was 2.690（0.717, 7.750） mg/L （z=-3.688, P〈0.05）. Meanwhile, the serum levels of PLA2R antibody in IMN patients were significantly different between different stages and ages （χ^2 values： 10.328, 9.716, both P〈0.05）. According to the receiver operating characteristic （ROC） curve, when the diagnostic cut-off was set at 0.80 mg/L for IMN detection, the sensitivity and specificity of anti-PLA2R-TRFIA were 73.0%（46/63） and 95.6%（86/90）, respectively.ConclusionsAnti-PLA2R-TRFIA has been well established. This quick and easy-performance method could increase the diagnostic accuracy for IMN.

关 键 词：肾小球肾炎 膜性 受体 磷酯酶A2 荧光免疫测定 

分 类 号：R446.6[医药卫生—诊断学]