摘要
目的建立一种检测溶血血清标本中血红蛋白浓度的自动化分析方法。方法利用血红蛋白具有类过氧化物酶活性的特性,根据四甲基联苯胺(TMB)的呈色原理,用以TMB为色原的酶联免疫吸附试验(ELISA)试剂盒的剩余A液(含过氧化氢)、B液(含TMB)作为显色剂,在一定范围内其颜色的深浅与血红蛋白的浓度呈正比,在660nm的波长下测定其吸光度的变化,进而计算出血红蛋白的浓度。并确定此法的精密度、线性范围、回收率及其干扰情况。结果该法测定3个不同溶血程度的血清标本的血红蛋白浓度,其批内平均CV值为1.96%,批间平均CV值为3.29%;当血红蛋白浓度为1～25g/L时,其相关系数(r)=0.999;3个不同溶血程度的血清标本其血红蛋白浓度的平均回收率为101.59%;终浓度为134μmol/L的胆红素标准液对3个样本的血红蛋白浓度检测的平均相对误差为2.34%,三酰甘油为5.08mmol/L的脂血标本对3个样本的血红蛋白浓度检测的平均相对误差为2.68%。结论本法重复性好,线性范围宽,结果准确,抗干扰能力强,可用于自动化分析,试剂无致癌作用、成本低、来源广,易于推广。
Objective To establish an automated analytical method for detecting the hemoglobin concentration in hemolytic serum specimens.Methods Using the enzymatic activity of hemoglobin as peroxidase,the color rendering principle of tetramethyl benzidine（TMB） as developer and the hemoglobin concentration with the color of the developer having direct proportion relationship in some limit,to determine the precision,linear range,recovery rate and interference.Results The average intraassay coefficient of variation in three samples with different hemolytic degrees was 1.96%,and the interassay coefficient was 3.29%.The coefficient correlation was 0.999 in the hemoglobin concentration range of 1-25 g/L.The average recovery rate of the hemoglobin concentration in three samples was 101.59%.The average relative error of the hemoglobin concentration was 2.34% when the bilirubin final concentration was 134 μmol/L,and that of the hemoglobin concentration was 2.68% when the triglyceride concentration was 5.08 mmol/L.Conclusion The method has good reproducibility,wide linear range,accurate result and good anti-interference ability,which can be used for automatic analysis.The reagent has noncarcinogenic action,low cost and broad source,which is easy to be widely spread.
出处
《检验医学与临床》
CAS
2010年第15期1537-1538,1541,共3页
Laboratory Medicine and Clinic
基金
佛山市卫生局资助项目(2008276)
关键词
血红蛋白
溶血
自动化
hemoglobin
haemolysis
automation
