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CysC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CysC抗血清包被乳胶颗粒,研制PE-TIA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8.11~0.25mg/L,检测限0.2mg/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.
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 self-preparing antibody to test human serum Cys C levels could be applied in automatic biochemical analyzer with satisfactory performance.
出处
《第四军医大学学报》
北大核心
2009年第21期2299-2301,共3页
Journal of the Fourth Military Medical University
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
cystatin C
antiserum
particle enhanced turbidimetric immunoassay
evaluation test of methodology