应用溶血指数探讨溶血对多个生化项目的干扰情况

目的:探讨标本溶血对生化项目检测结果的影响,并使用溶血指数对影响程度进行量化,并探讨其临床意义。方法:总蛋白(TP)、尿酸(UA)、肌酸激酶(CK)、乳酸脱氢酶(LDH)、总胆红素(TBIL)均参考美国临床实验室标准化委员会(CLSI)制定的评价方案EP7-A2文件的标准程序,每个项目收集20份新鲜混合血清(TBIL<17.1μmol/L和TBIL>150μmol/L分别收集),包括参考值范围内的10份、低值和高值各5份,每份新鲜混合血清用3~4份入选血样混合成约8 mL。每份新鲜混合血清分成若干份,分别与洗涤好红细胞配制的溶血液制成5个不同浓度的溶血模型(试验组)及与之配对的无溶血模型(对照组),然后分别测定试验组、对照组及分出的1份混合血清原样的血红蛋白浓度,并在罗氏cobasC501全自动生化分析仪上测定3次所需项目和溶血指数,计算血红蛋白浓度与溶血指数之间的关系,以及溶血对TP、UA、CK、LDH、TBIL检测结果影响的大小。参考EP7-A2文件的统计方法,计算最大允许偏差对应的干扰物浓度,以每份测试混合血清的溶血指数为x轴、其对应的检测项目变化值(干扰值)为y轴绘制干扰曲线。结果:对每项目20份混合血清的660个测试、5个项目3960检测结果用假设检验统计,TP、CK、LDH、TBIL(<17.1μmol/L和>150μmol/L)的μ值分别为4.42、2.34、2.20、2.76、2.93,P值均小于0.05,说明溶血对TP、UA、CK、LDH、TBIL均存在显著干扰。通过回归分析统计出不同溶血程度的溶血指数值对应的干扰变化值的干扰剂量反应曲线。TP、CK、LDH、TBIL的干扰剂量回归方程分别为y=0.0099x-0.065、y=0.1001x-2.42、y=1.1406x-1.3303、y=1.78×10-5x2-0.011x-0.235、y=1.88×10-5x2-0.048x-4.169。从干扰剂量反应曲线可看出溶血对LDH、CK、TP检测产生正干扰,对TBIL检测产生负干扰。同时发现溶血对UA结果的干扰很不稳定,即溶血对尿酸检测结果存在正干扰及负干扰,这一结果得到相关文献印证。结论:溶血指数可作为标本是否合格的客观指标,避免主观判断的误差;利用测得的溶血指数,可从干扰剂量曲线或回归方程求出检测结果的变化值,使临床通过干扰曲线判断溶血对检测项目产生的影响,从而做出准确客观的判断。

Application of Hemolytic Index in the Project of Hemolysis Interfer ence on a Number of Biochemical Project

Objective: To investigate the infection of hemolysis sample in the results of biochemical test,and to quantify the influence degree of hemolysis index,and to discuss the clinical significance.Methods: According to the standard procedure files EP7-A2 established in the Clinical Laboratory Standardization Commission(CLSI) of USA,we made a project to detect the relationship between hemoglobin concentrations and hemolysis index,and the hemolysis influence to the total protein(TP),uric acid(UA),creatine kinase(CK) and lactate dehydrogenase(LDH),total bilirubin(TBIL).We taked the fresh,no hemolysis,no jaundice and no fat serum from 10 cases with in the reference,5 cases of low value and 5 cases of high value,and we made some washing erythrocyte to exe cute 5 different concentrations hemolysis model as experiment group and paired with no hemolysis model as con trol group.Hemoglobin concentrations were detected in the two groups and the original mixture serum,the whole items and hemolysis index were detected for three times in ROCH automatic biochemistry analyzer.We made a disturbing curve for the x-axis of the hemolysis index of mixed serum and y-axis of the corresponding detection program changes(interference).Refered to the statistical methods of EP7-A2,we work out the disturbing concentra tion corresponding the maximum allowable deviation.Results: The statistical results showed that μ value of TP,CK,LDH,TBIL(<17.1 μmol/L and >150 μmol/L) were 4.42,2.34,2.20,2.76 and 2.93 due to 660 tests and 3960 detection results in 20 mixture serum,P value were all less than 0.05 means hemolysis obviously disturb the TP、CK、LDH and TBIL.The interference dose response curve was made through the statistics regression analy sis.The interference doses regression equation of TP,CK,LDH,TBIL were y =0.0099 x-0.065,y =0.1001 x-2.42,y = 1.1406 x-1.3303,y =1.78×10-5 x 2-0.011 x-0.235,y =1.88×10-5 x 2-0.048 x-4.169.According to the interference dose re sponse curve,we get that hemolysis produced positive interference to the results of LDH,CK,TP,and reverse in terference to the results of TBIL.At the same time,we found that the hemolysis interference was not stable to the UA,which means hemolysis can produce both positive and reverse interference,and consult literature also con firmed that.Conclusion: The hemolysis index can be used as objective indicators of specimens eligibility and avoid subjective judgment error.On the measurement of hemolysis index and test results of a value change,inter ference dose response curve was made to judge hemolysis interference of the biochemical item detection,thus make accurate objective judgment.