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[1]吴祝圆,黄毅文,贾子超,等.尿游离皮质醇萃取方法的改良及其检测相关影响因素的分析[J].天津医科大学学报,2022,28(04):436-441.
 WU Zhu-yuan,HUANG Yi-wen,JIA Zi-chao,et al.Improvement of extraction method and analysis of the interferences in the detection of urinary free cortisol[J].Journal of Tianjin Medical University,2022,28(04):436-441.
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尿游离皮质醇萃取方法的改良及其检测相关影响因素的分析(PDF)
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《天津医科大学学报》[ISSN:1006-8147/CN:12-1259/R]

卷:
28卷
期数:
2022年04期
页码:
436-441
栏目:
技术与方法
出版日期:
2022-07-20

文章信息/Info

Title:
Improvement of extraction method and analysis of the interferences in the detection of urinary free cortisol
文章编号:
1006-8147(2022)04-0436-06
作者:
吴祝圆1黄毅文1贾子超1胡志东1罗微12
(1.天津医科大学总医院检验科,天津 300052,2.天津医科大学医学技术学院,天津 300023)
Author(s):
WU Zhu-yuan1HUANG Yi-wen1JIA Zi-chao1HU Zhi-dong1LUO Wei12
(1.Department of Clinical Laboratory,General Hospital,Tianjin Medical University,Tianjin 300052,China;2.School of Medical Laboratory,Tianjin Medical University,Tianjin 300203,China )
关键词:
尿游离皮质醇萃取改良二氯甲烷影响因素
Keywords:
urinary freecortisolextractionimprovementdichloromethane influencing factors
分类号:
R331.1
DOI:
-
文献标志码:
A
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=0.998 9,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R2=0.992 2。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。
Abstract:
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 23 24-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 0.998 9. 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 0.992 2. 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 decrease the values.

参考文献/References:

[1] CECCATO F,BOSCARO M. Cushing′s syndrome:screening and diagnosis[J]. High Blood Press Cardiovasc Prev,2016,23(3):209-215.
[2] LUO A,EL GIERARI ETM,Nally L M,et al. Clinical utility of an ultrasensitive urinary free cortisol assay by tandem mass spectrometry[J]. Steroids,2019,146:65-69.
[3] ALEXANDRAKI K I,GROSSMAN A B. Is urinary free cortisol of value in the diagnosis of Cushing′s syndrome? [J]. Curr Opin Endocrinol Diabetes Obes,2011,18(4):259-263.
[4] YATSCOFF R W,CHAPELSKY L,MORRISH D. Analytical and clinical evaluation of an automated cortisol assay on the ACS:180[J]. Clin Biochem,1996,29(4):315-319.
[5] KOWALSKI A,PAUL W. A simple extraction procedure for the determination of free (unconjugated) cortisol in urine by radioimmunoassay[J]. Clin Chem,1976,25:1152-1158.
[6] 林瑶,黄志基,胡尧,等. 电化学发光免疫分析24小时尿游离皮质醇的方法学验证[J]. 标记免疫分析与临床,2014,21(3):306-309.
[7] 熊中云,马怡,施绍瑞,等. 盐酸对24小时尿游离皮质醇测定的影响[J]. 华西医学,2010,25(6):1114-1116.
[8] 张殿喜,李乃适,黎明,等. 影响用化学发光免疫法测定尿游离皮质醇的重要因素[J]. 中国卫生检验杂志,2009,19(7):1564-1565.

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备注/Memo

备注/Memo:
基金项目 国家自然科学基金(81701968)
作者简介:吴祝圆(1990-),女,硕士在读,研究方向:内分泌免疫诊断;通信作者:胡志东,E-mail:huzhidong27@163.com;罗微,E-mail: weiluo1028@tmu.edu.cn。
更新日期/Last Update: 2022-07-20