尿清蛋白、β_2微球蛋白的双标记TrFIA法的建立  

Establishment and evaluation of a dual-labeled time-resolved fluoroimmunoassay for simultaneously detecting β2-MG and albumin in urine

作　　者：马智鸿[1] 黄飚[1] 张珏[1] 张艺[1] 刘海燕[1] 

机构地区：[1]江苏省原子医学研究所,卫生部核医学重点试验室,江苏无锡214063

出　　处：《临床检验杂志》2009年第5期360-362,共3页Chinese Journal of Clinical Laboratory Science

摘　　要：目的建立同时可测尿清蛋白(Alb)和β2微球蛋白(β2-MG)的双标记时间分辨荧光免疫分析法(β2-MG/AlbTr FIA)。方法用羊抗兔IgG包被96孔板,加入限量的特异性兔抗人Alb抗体和兔抗人β2-MG抗体使之结合到板上,使Eu3+标记的β2-MG和Sm3+标记的Alb与标准品(标本)中的β2-MG、Alb竞争结合在板上的限量相应抗体,以Log-Logit制作标准曲线,并对方法学进行评价。结果该法检测尿液Alb和β2-MG的灵敏度分别为0.2mg/L和0.05mg/L。检测范围Alb为2～50mg/L,β2-MG为0.1～8mg/L。检测Alb平均批内、批间CV%分别为4.6%、7.8%;检测β2-MG的平均批内、批间CV%分别为3.3%、5.5%。Alb和β2-MG的平均添加回收率分别为95.4%和102.4%。该法结果与单独检测Alb和β2-MG的放射免疫方法所测结果配对t检验均P>0.05。结论该方法可同时定量检测尿Alb和β2-MG,简便、快速,结果准确,适于临床应用。Objective To develop a dual-labeled time-resolved fluoroimmunoassay （TRFIA） to simultaneously determine albumin （Alb） and β2 -microglobulin （ β2 -MG） in urine. Methods The 96 wells were coated with goat anti-rabbit IgG, followed by adding limited rabbit anti-Alb IgG and rabbit anti-β2-MG IgG to bind to the coated IgG. Sm^3＋ labeled Alb and Eu^3＋ labeled β2-MG were added to compete with Alb and β2-MG in the standards/samples respectively. The standard curves were established by log-logit function by the DELFIA instrument. The precision, accuracy, and sensitivity of the assay were evaluated. Results The low detection limit was 0.2 mg,/L for Alb and 0.05 mg/L for β2-MG. The detection ranges for Alb and β2-MG were 2 - 50 mg/L and 0.1 - 8 mg/L respectively. The within-run and between-run coefficient of variation values for Alb were 4.6% and 7.8% ,and for β2-MG were 3.3% and 5.5% respectively. The results obtained by the assay showed good correlation with those obtained by RIA kit. Conclusions The β2-MG/Alb TRFIA developed in the present study may simultaneously determine the Alb and β2-MG in the urine. The assay is simple,rapid, and accurate in the detection, and is suitable for the clinical application.

关 键 词：双标记 时间分辨荧光免疫分析 尿液 清蛋白 Β2微球蛋白 肾脏病 

分 类 号：R446.6[医药卫生—诊断学]