自制抗体的颗粒增强透射免疫分析法测定人血清胱抑素C的评价
Evaluation of particle enhanced turbidimetric immunoassay method for human serum Cys C with selfpreparing antibody
陈婷梅 1俸家富 2郭变琴 1文阳安 1涂植光1
作者信息
- 1. 重庆医科大学检验系,教育部临床检验诊断学重点实验室,重庆,400016
- 2. 绵阳市中心医院检验科,四川,绵阳,621000
- 折叠
摘要
目的:用自制抗体研制可在自动生化分析仪应用的颗粒增强透射免疫浊度分析法(PETIA)定量测定人血清胱抑素C(Cys C)的试剂盒.方法:构建人Cys C的原核表达载体pET32a(+)/Cys C,纯化Cys C重组蛋白并免疫大白兔,制备Cys C抗血清;以自制的Cys C抗血清包被乳胶颗粒,研制PETIA法定量检测血清Cys C的试剂盒,并对其分析性能进行方法学评价.结果:本法回收率为97.5%～105.0%,批内及批间CV均＜5%,线性范围8.11～0.25 mg/L,检测限0.2 mg/L;本法与Dako法和BN-Prospect法呈正相关(P＜0.0001);胆红素对高浓度Cys C测定有干扰作用,Hb和葡萄糖对本法无干扰.结论:应用自动生化分析仪的自研法性能满意.
Abstract
AIM:To develop a kit of particle enhanced turbidimetric immunoassay(PETIA)method based on self-made antibody for determining human serum Cys C level with automatic biochemical analyzer.METHODS:The prokaryotic expression vector pET32a(+)/Cys C was constructed,in which Cys C expression was induced.The antiserum was prepared after rabbits were immunized with the purified recombinant protein.Then the antibody was covalently attached to the uniform microparticles,a kit of PETIA method for human serum Cys C test was established,and methodology evaluations were done.RESULTS:The recovery rate was 97.5%-105.0%,CV was less than 5%,linear range 8.11-0.25 mg/L,minimum limit 0.2 mg/L.The performance of self-established PETIA was equal to the import kits(Dako & BN-Prospect).The bilirubin had an effect on assays of high level Cys C,while Hb and glucose had no interference.CONCLUSION:The kit of PETIA based on serf-preparing antibody to test human serum Cys C levels could be applied in automatic biochemical analyzer with satisfactory performance.
关键词
胱抑素C/抗血清/颗粒增强透射免疫浊度分析法/方法学评价Key words
cystatin C/antiserum/particle enhanced turbidimetric immunoassay/evaluation test of methodology引用本文复制引用
出版年
2009
NETL
NSTL
维普
万方数据