免疫比浊法测定尿液游离血红蛋白的方法学研究  

Methodological Research on Latex Enhanced Immune Turbidimetric Method for the Detection of Urine Free Hemoglobin

作　　者：朱海波[1,2] 尹伶[2] 王新[1] 严念道[1] 邓建平 ZHU Hai-bo;YIN Ling;WANG Xin;YAN Nian-dao;DENG Jian-ping(Department of Clinical Laboratory ,Aikang Hospital, Hubei Huangshi 435000, China;Wuhan Medical School of Science and Technology University ,Wuhan 430081 ,China)

机构地区：[1]黄石市爱康医院检验科,湖北黄石435000 [2]武汉科技大学医学院,武汉430081

出　　处：《现代检验医学杂志》2018年第1期67-71,76,共6页Journal of Modern Laboratory Medicine

基　　金：湖北省黄石市市级科研项目(黄卫生计生通[2014]72)

摘　　要：目的在全自动生化分析仪上建立胶乳增强免疫比浊法测定尿液游离血红蛋白的方法学,并进行方法性能评价。方法使用抗人血红蛋白HBA0抗体(8.5mg/ml),80nm聚苯乙烯羧基胶乳微球(estapor,纳米微球)等材料制备检测试剂;在东芝TBA120全自动生化仪上设置项目参数并进行校准;方法学性能评价包括准确度、精密度、线性范围、灵敏度、特异度等性能指标;建立该地区健康人群尿液游离血红蛋白的生物参考区间并验证。结果项目参数建立后定标通过,各测定点与拟合的校准曲线未呈现明显偏离,定标液浓度值与吸光度值经相关性分析,r=0.980 7,r^2=0.961 8;回收试验中低值样本S_低加入浓度为100mg/L和500mg/L的标准液后回收率分别为95.3%和102.7%,高值样本S_高加入浓度为100mg/L和500mg/L的标准液后回收率分别为104.2%和103.5%;精密度试验中低值样本S_低和高值样本S_高的总不精密度CV分别的6.52%和4.18%;线性范围为0~1 100mg/L;分析灵敏度为2.8mg/L;干扰试验显示当样本中游离胆红素、结合胆红素、维生素C、动物血红蛋白分别小于342μmol/L,342μmol/L,0.03g/L和500mg/L时对该试验结果无明显干扰,当乳糜浓度>870FTU时对该试验结果有显著干扰;该研究建立的健康人群生物参考区间男性为0~13.3mg/L、女性为0~17.1mg/L,两组参考区间经t检验无显著性差异(P>0.05)。结论研究表明该方法学性能具备检测临床尿液标本,解决了化学法潜血试验特异性不够的问题,并实现了数值化检测,该研究也为其他类型临床标本的检测提供了思路。Objective To establish the test way of latex enhanced immune turbidimetric method for the concentration detection of urine free hemoglobin on the automatic biochemical analyzer,and evaluated the detection performance.Methods Prepared the detection reagents of latex enhanced immune turbidimetry method for detection of urine free hemoglobin with materials such as hemoglobin HBA0 antibody（8.5 mg/ml）,80 nm carboxylic olystyrene latex microspheres（estapor,nanometer microspheres）and so on.Then,seted the project parameters on TBA120 automatic biochemical analyzer and proceeded the calibrations.Evaluated the detection performance by accuracy,precision,linear range,sensitivity,specificity and other indicators.Lastly,established the biological reference interval for urine free hemoglobin of healthy people in the region and verified it.Results To seted the project parameters on TBA120 automatic biochemical analyzer and passed the calibration.The calibration fitting curve had a smooth trend and no obvious deviation with each measuring point.In the recovery tests,added standard liquid with different concentrations of 100 mg/L and 500 mg/L to the samples with low values,the recovery rate was 95.3% and 102.7%respectively.Same to the samples with high values,the recovery rate was 104.2% and 103.5%respectively.In the precision test,samples with low and high value had a precision CVof 6.52% and 4.18%respectively.The linear range was 0-1 100 mg/L,analysis sensitivity was 2.8 mg/L.In the interference test,found that,samples had no obvious disturbance to the test with lower concentrations of free bilirubin,combined with bilirubin,vitamin C,animal hemoglobin lower than 342μmol/L,342μmol/L,0.03 g/L,500 mg/L respectively,while had significant interference when the concentration of chyle was higher than 870 FTU.Their study had established the biological reference interval of healthy people,male was 0-13.3 mg/L and female 0-17.1 mg/L,there was no significant difference between them by at test（P〈0.05）.Conclusion The latex e

关 键 词：胶乳 免疫比浊法 尿液 游离血红蛋白 方法学 

分 类 号：R446.121[医药卫生—诊断学]