摘要
目的:改良24 h尿游离皮质醇(urinary free cortisol,UFC)的萃取方法,分析标本留置时间、蛋白尿及潜血尿对结果的影响。方法:取2019年12月—2020年12月天津医科大学总医院检验科的23例不同结果的24 hUFC标本,其中8例低值、8例中值、7例高值。采用二甲氯烷萃取-化学发光法检测UFC。在萃取步骤中用恒速回旋振荡(改良法)替代触碰式涡旋振荡(原方法);改良后设置不同标本留置时间,配制蛋白及潜血尿,检测UFC,分析这些干扰因素对检测的影响。结果:改良法与原方法对比,效率更高且萃取成功率提升至100%,两种方法检测结果无差异(P>0.05)且相关系数R2=0.9989,改良法能提升检测精密度[变异系数(CV)<25%]与准确度(>80%);与液相色谱-串联质谱法(LC-MS/MS)相比差异无统计学意义(P>0.05),相关系数R^(2)=0.9922。尿标本不同留置时间对结果影响差异无统计学意义(P>0.05),尿蛋白及潜血干扰实验的平均回收率分别为95.85%及89.85%,但尿血红蛋白(Hb)达4 mg/L时会导致UFC检测误差大于该项目的实验允许总误差(TEA,25%)。结论:恒速回旋振荡萃取可替代触碰式涡旋振荡萃取法,可提高工作效率且不影响检测结果。标本留置时间、蛋白尿对检测结果影响不大,但尿潜血会使检测值偏低。
Objective:To improve the extraction method and analyze the interferences such as the sample retention time,urine protein,occult blood and urine output in the detection of 24 h urinary free cortisol(UFC).Methods:A total of 2324-hour UFC samples with different results from the Department of Clinical Laboratory of the General Hospital of Tianjin Medical University were collected.Samples with low(n=8),medium(n=8)and high(n=7)UFC concentration in original method were selected,and redetected by modified dichloromethane extraction method with chemiluminescence immunoassay.At the same time,vortex oscillator with contact type in original extraction method was changed into shaker oscillator with a constant-speed cyclotron oscillating(improved method).After changing to the improved method,UFC with different interferences such as storage time,urine protein,occult blood and urine output were detected after artificial setting.Results:Compared with the original method,the improved constant speed extraction method improved the extraction success rate,and there was no statistical difference in the detection results(P>0.05)and the correlation coefficient R2 was about 0.9989.The improved method couldimprove detection precision(coefficient of variation<25%)and accuracy(>80%).Compared with liquid chromatography-tandem mass spectrometry(LC-MS/MS),there was also no difference(P>0.05)and the correlation coefficient R2 was about 0.9922.Different retention time of urine specimens had no effect on the results(P>0.05).The average recovery of urine protein and urine occult blood interference experiments were 95.85%,89.85%,respectively.But when urine hemoglobin(Hb)reached 4 mg/L,the error would be greater than the total error allowable(TEA,25%)of UFC detection.Conclusion:The improved shaker constant speed oscillation extraction method can replace the original touch vortex oscillation method and does not affect the detection results.Urine specimen retention time and proteinuria have little effect on the test results,but occult blood urine will
作者
吴祝圆
黄毅文
贾子超
胡志东
罗微
WU Zhu-yuan;HUANG Yi-wen;JIA Zi-chao;HU Zhi-dong;LUO Wei(Department of Clinical Laboratory,General Hospital,Tianjin Medical University,Tianjin 300052,China;School of Medical Laboratory,Tianjin Medical University,Tianjin 300203,China)
出处
《天津医科大学学报》
2022年第4期436-441,共6页
Journal of Tianjin Medical University
基金
国家自然科学基金(81701968)。
关键词
尿游离皮质醇
萃取
改良
二氯甲烷
影响因素
urinary freecortisol
extraction
improvement
dichloromethane
influencing factors
