活性氧代谢物衍生物和生物抗氧化能力检测方法的建立及其在评价甲状腺功能亢进疾病中的作用

发布时间：2018-01-05 23:28

本文关键词：活性氧代谢物衍生物和生物抗氧化能力检测方法的建立及其在评价甲状腺功能亢进疾病中的作用　出处：《北京协和医学院》2015年硕士论文　论文类型：学位论文

【摘要】：目的基于全自动生化分析仪,建立检测血清中氧化代谢物衍生物(Derivatives of Reactive Oxygen Metabolites, DROM)和生物抗氧化能力(Biological Antioxidant Potential, BAP)的检测方法,并对其检测性能进行评估,建立健康人参考区间。比较健康对照与甲状腺功能亢进患者血清DROM和BAP水平差异,通过研究DROM和BAP水平与甲状腺功能亢进疾病之间的相关性,探究氧化应激指标DROM和还原指标BAP在评价甲亢中的作用方法根据DROM和BAP的检测原理,建立可应用于全自动生化分析仪的氧化还原体系血清学指标。参考美国国家临床和实验室标准协会(Clinical and Laboratory Standards Insitute, CLSI) EP15-A2、EP17-A、EP6-A、EP7-P方案对该方法的精密度、空白限(LoB)、检出限(LoD)、定量检出限(LoQ)、线性范围、干扰因素进行评估,以此分析新建立的DROM和BAP检测方法的检测性能。分别选取健康非吸烟成年人300例和287例进行DROM和BAP检测,建立正常人参考区间。2014年11月1日至2015年3月31日于卫生部北京医院门诊治疗的甲状腺亢进患者73例,选取年龄性别匹配的健康人100例。所有受试者都进行了DROM、 BAP、SOD氧化应激相关指标的检测以及TSH、FT3、FT4甲状腺相关激素的检测。结果1) DROM批内不精密度为2.38%—3.85%,总不精密度为4.33%—6.67%;空白限LoB为2.85mmol/L过氧化氢叔丁醇,检出限LoD为11.43mmol/L过氧化氢叔丁醇,定量检出限LoQ为26.88mmol/L过氧化氢叔丁醇；线性范围为31.77-400.00mmol/L (r2= 0.995);三种干扰物在试验浓度内(甘油三脂≤27.18mmol/L, Hb175g/L,抗坏血酸60mg/d1)引起的偏差分别为-5.62%,309.57%及36.6%;正常参考区间为≤107 mmol过氧化氢叔丁醇。2)BAP全自动生化分析仪的低浓度标准品总不精密度为5.87%,高浓度标准品总不精密度为5.80%,低浓度血清总不精密度为14.14%,高浓度血清总不精密度为12.61%。空白限LoB= 0.068 mmol/L VitC,血清标本检测限LoD=0.154mmol/L VitC,定量检测限LoQ=0.224 mmol/L VitC.线性范围为0-4 mmol/L VitC(r2=0.9996)。三种干扰物在试验浓度内(血红蛋白≤170g/L,甘油三酯7.385mmol/L,胆红素≤73.8 μmol/L)的偏差分别为-2.58%、-13.11%、83.1%。健康生物参考区间为≥2.89 mmol/L VitC当量。3)入组的甲亢病人与健康对照之间DROM, BAP之间有明显差异,但是DROM与TSH、FT3、FT4都没有明显相关性;BAP与TSH、FT3、FT4没有明显相关性；SOD与TSH呈正相关,SOD与FT3呈负相关,SOD与FT4没有明显相关性。结论新建立的DROM和BAP检测方法具有操作简便、检测速度快、高通量自动检测等优点,适用于临床实验室普遍使用的全自动生化分析仪。该方法具有良好的精密度、线性范围和抗干扰能力,可满足临床检测要求,DROM正常人参考区间为≤107 mmol过氧化氢叔丁醇,BAP正常人参考区间为≥2.89 mmol VitC当量。DROM和BAP在甲亢病人的病情评估中有临床意义。
[Abstract]:Based on the automatic biochemical analyzer, establish serum oxidative metabolites derivatives (Derivatives of Reactive Oxygen Metabolites, DROM) and biological antioxidant capacity (Biological Antioxidant Potential, BAP) detection method, and evaluate the detection performance, establish healthy reference interval. Differences between healthy controls and patients with hyperthyroidism and serum DROM the level of BAP, through the study of the correlation between the DROM and BAP levels with hyperthyroid disease, explore the oxidative stress markers DROM and BAP index in the evaluation of a reduction in the role of Kang methods according to the detection principle of DROM and BAP, can be used to establish the oxidation reduction system of automatic biochemical analyzer serological indicators refer to national clinical and laboratory. Standard Association (Clinical and Laboratory Standards Insitute, CLSI) EP15-A2, EP17-A, EP6-A, EP7-P The precision of the method, limit of blank (LoB), the limit of detection (LoD), quantitative detection limit (LoQ), linear range, interference factors evaluation, detection performance analysis of DROM and BAP on the detection of the new method. Selected healthy non-smoking adults in 300 cases and 287 cases in DROM and BAP detection, establish a normal reference range of.2014 in November 1st to March 31, 2015 in Beijing Hospital of Ministry of health outpatient treatment of hyperthyroidism patients in 73 cases, healthy people age and gender matched 100 cases. All of the subjects were DROM, BAP, SOD, oxidative stress related indicators and detection of TSH, FT3, FT4 detection of thyroid related hormones the results of DROM. 1) within run precision was 2.38% - 3.85%, total imprecision is 4.33% - 6.67%; LoB is the limit of blank 2.85mmol/L hydrogen peroxide TERT butanol, the detection limit of LoD 11.43mmol/L hydrogen peroxide, tert butanol, quantitative detection limit is 26.88mmol/ LoQ L hydrogen peroxide tert butyl alcohol; the linear range of 31.77-400.00mmol/L (r2= 0.995); three types of distractors in the test concentration in (glycerin three greases = 27.18mmol/L, Hb175g/L, ascorbic acid 60mg/d1) caused by the deviation of -5.62% respectively, 309.57% and 36.6%; normal reference interval is less than or equal to 107 mmol hydrogen peroxide tert butyl.2) low concentration standard BAP automatic biochemical analyzer of total imprecision was 5.87%, high concentration of standard total imprecision was 5.80%, low concentration of serum total imprecision was 14.14%, high concentration of serum total imprecision of 12.61%. LoB= 0.068 mmol/ L VitC blank serum samples, the detection limit of LoD=0.154mmol/L VitC, LoQ=0.224 mmol/L VitC. quantitative detection limit of the linear range for 0-4 mmol/L VitC (r2=0.9996). Three types of distractors in the test concentration within (hemoglobin less than 170g/L, triglyceride 7.385mmol/L, bilirubin is less than or equal to 73.8 mu mol/L) deviation were -2.58%, -13.11% 83.1%. health, biological reference interval of not less than 2.89 mmol/L VitC equivalent of.3) in the group of patients with hyperthyroidism and healthy controls DROM, there was significant difference between BAP and TSH, but DROM, FT3, FT4 have no obvious correlation; BAP and TSH, FT3, FT4 have no significant correlation; a positive correlation between SOD and TSH, negative correlation between SOD and FT3, there was no significant correlation between SOD and FT4. DROM and BAP detection method of the new conclusion has the advantages of simple operation, fast detection speed, high throughput detection etc., application of automatic biochemical analyzer is widely used in clinical laboratories. This method has good precision, linear range and anti-jamming ability. Can meet the clinical detection requirements, DROM normal reference interval is less than or equal to 107 mmol hydrogen peroxide, tert butanol, BAP normal reference range for more than 2.89 mmol VitC has clinical significance in evaluation of.DROM and BAP in patients with hyperthyroidism equivalent condition.

【学位授予单位】：北京协和医学院
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R446.1;R581.1

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