颗粒增强免疫透射比浊测定血中Cyst C全自动分析法的建立与评价  

Appraisal of a New Method of Automated Particle-Enhanced Turbidimetric Immunoassay for the Routine Measurement of Human Cystatin C Established in Our Laboratory

作　　者：贾宁人[1] 吴家明[2] 芮志莲 李康[3] 王宁皎[1] 蒋维[1] 

机构地区：[1]江苏省中医院检验科 [2]溧阳市人民医院 [3]东南大学

出　　处：《放射免疫学杂志》2006年第5期423-426,共4页Journal of Radioimmanology

基　　金：2004年常州市卫生局科技项目

摘　　要：目的:建立颗粒增强免疫透射比浊测定血中胱蛋白酶抑制剂C(Cyst C)全自动分析,评价该法常规用于检测血中胱蛋白酶抑制剂C的可行性。方法:将含有Cyst C血样本与乳胶颗粒增强的Cyst C多克隆兔抗体按一定体积比混合,在一定量的兔免疫球蛋白及表面活性剂存在条件下,抗原、抗体结合反应在一定温度、时间内产生一定大小的颗粒,许多颗粒在反应液体中形成一定的浊度,利用Olympus AU2700全自动生化分析仪透射比浊测定,并对方法的不精密度、准确度、干扰试验及与Dade Behring BN ProSpec免疫散射比浊测定方法比较等进行评价。结果:Cyst C浓度为1.03mg/L、5.67mg/L样本的批内不精密度CV分别为4.8%、1.9%。向Cyst C浓度为0.79mg/L、1.18mg/L、1.46mg/L、2.46mg/L样本中加入1.65mg/L的标准液测定回收率分别为100%、95%、85%、87%,平均回收率为92%。样本中的类风湿因子、胆红素、血红蛋白、三酰甘油等干扰物浓度分别达975U/ml、400μmol/L、5g/L、6.7mmol/L时对检测无显著性影响。Cyst C浓度在0.41mg/L^6.60mg/L范围内检测线性良好。血清、肝素抗凝血浆及EDTA抗凝血浆样本测得的Cyst C结果未发现显著性差异(F=0.065,P=0.938)。与Dade Behring BN ProSpec免疫散射比浊测定方法比较有良好的相关性,Y=0.98x-0.054,r=0.997,P<0.01,但两者存在非常显著性差异(P<0.01),本法结果较Dade Behring BN ProSpec免疫散射仪测定结果略低,两种方法的平均偏差为-0.0678mg/L。结论:建立的颗粒增强免疫透射比浊法测定血CystC重复性好、准确度高。血清、肝素抗凝血浆及EDTA抗凝血浆均可用于分析。测定不受类风湿因子(≥975U/ml),胆红素(≥400μmol/L),血红蛋白(≥5g/L)及三酰甘油(≥6.7mmol/L)的干扰。与Dade Behring BNProSpec免疫散射比浊测定方法测定结果比较有良好的相关性,适用于在全自动生化分析仪上作常规检测。Objective To establish a new method of automated particle - enhanced turbidimetric immunoassay for the serum cystatin C and evaluate its feasibility for routine measurement. Methods The tested serum sample containing cystatin C was mixed with polyclonal rabbit anti-human cystatin C antibody. In the presence of rabbit immunoglobulin and surfactant, the binding reaction between antigen and antibody under optimal temperature and time would complete and produce particles, causing turbidity in the reaction fluid. We determined the turbidity of the reaction fluid with turbidimitric method （PETIA） using Olympus AU2700 automated biochemistry analyzer, and examined the imprecision, accuracy, interference test of the method. We also compared the result from this method with that from the Dade Behring BN ProSpec immunonephelometric assay. Results The intra CV imprecision of the samples that contain 1.03mg/L and 5.67mg/L cystatin C was 4.8% and 1.9% respectively. Adding 1.65mg/L standard solution into samples with cystatin C concentration of 0.79mg/L, 1.18mg/L, 1.46mg/L, 2.46mg/L respectively, and we got their recovery rate as 100%, 95%, 85%, 87% respectively （mean 92%）. Rheumatoid factor （≥975U/ml）, bilirubin （≥400μmol/L）, haemoglobin （≥5g/L）, and triglycerides （≥6.7mmol/L） in the tested samples had no significant interfernce in the assay. When the cystatin C concentration was between 0.41mg/L～6.60mg/L, the method showed good linearity. The method could be applied to serum, heparin - plasma and EDTA - plasma specimens with similiar results. Resfult of this method correlated well （Y = 0.987x - 0.054, r = 0.997） with that of the Dade Behring BN ProSpec particle - enhanced immunonephelometric assay （PEINA）, but there were significant differences （P 〈 0.01）. Value from this method were slightly lower than that from the Dade Behring BN ProSpec particle - enhanced immunonephelometric assay, the mean deviation was -0.0678mg/L. Conclusion This atuomated particle - enhanced turbidimet

关 键 词：胱蛋白酶抑制剂C(Cyst C) 颗粒增强的免疫透射比浊法(PETIA) 颗粒增强的免疫散射比浊法(PEINA) OLYMPUS AU2700全自动生化分析仪 肾小球滤过率(GFR) 

分 类 号：R446.112[医药卫生—诊断学]