液相色谱-质谱法、酶联免疫法和电化学发光法测定血清25-羟维生素D方法学比较
发布时间：2018-12-14 02:05
【摘要】：目的:比较国内三种主流方法:液相色谱-质谱法(LC-MS/MS)、电化学发光法(ECLIA)以及酶联免疫法(ELISA),同时测定同批血清25-羟维生素D(25(OH)D)的差异性、一致性。方法:于2015-07-01至2015-07-07,到河北医科大学第二医院儿科内分泌门诊行健康体检的儿童,共64份高低不同维生素D状况的血清。每份血清同时应用LC-MS/MS测定25(OH)D2、25(OH)D3浓度、ECLIA测定总25(OH)D浓度及ELISA(IDS)测定总25(OH)D浓度,进而应用SPSS17.0软件对每两种方法测定结果进行比较、直线相关关系分析、Bland-Altman法一致性检验。结果:1 LC-MS/MS测定64例血清25(OH)D数据符合正态分布,均数为65.37nmol/L,标准差为15.73nmol/L;ELISA和ECLIA测定结果均为非正态分布,中位数分别为:61.92nmol/L、60.18 nmol/L。Wilcoxon检验示每两种方法之间均数(中位数)差异均无统计学意义。箱式图示三种方法测定结果的平均水平均接近;ELISA与LC-MS/MS测定结果整体分布较一致,而ECLIA与其他两种方法相比,整体数据分布存在较大差异;2若将25(OH)D浓度50nmol/L、50-72.5nmol/L分别定义为维生素D缺乏、不足的切值,64例儿童中维生素D缺乏症的比例分别是:15.63%(ELISA)、17.18%(LC-MS/MS)、31.25%(ECLIA);维生素D不足的比例分别是:57.82%(ELISA)、54.69%(LC-MS/MS)、39.06%(ECLIA)。ELISA和LC-MS/MS结果一致,ECLIA与前两者差异较大;3每两种方法之间线性回归方程及相关系数为:Y(ECLIA)=1.32X(LC-MS/MS)-22.2(r=0.93);Y(ECLIA)=1.05X(ELISA)-4.73(r=0.81;Y(ELISA)=0.91X(LC-MS/MS)+5.79(r=0.84)。三种方法间均具有高度正相关性;4 ECLIA与LC-MS/MS的Bland-Altman图:4/64的点位于95%一致性界限外,在一致性界限内两方法相比,差值绝对值最大为17.05nmol/L,比值最大、最小分别为1.3、0.65;ELISA与LC-MS/MS:4/64的点位于95%一致性界限外,一致性界限内差值绝对值最大18.17nmol/L,比值最大、最小分别为1.3、0.75;ECLIA与ELISA:3/64的点位于95%一致性界限外,界限内差值绝对值最大为29.42nom/L,比值最大、最小分别为1.5、0.6。结论:当血清总25(OH)D浓度在一定范围内,且25(OH)D2含量极低时,ECLIA、LC-MS/MS、ELISA测定血清25(OH)D具有高度正向相关性,结合箱式图、直线回归方程、一致性检验分析,ECLIA所测血清25(OH)D水平与LC-MS/MS、ELISA具有一定差异性,而LC-MS/MS、ELISA差异性小、一致性佳。
[Abstract]:Objective: to compare the differences of simultaneous determination of 25 hydroxyvitamin D (25 (OH) D) in serum with three main methods: liquid chromatography-mass spectrometry (LC-MS/MS), electrochemiluminescence (ECLIA) and enzyme-linked immunosorbent assay (ELISA),). Consistency. Methods: 64 children with different levels of vitamin D were enrolled in the pediatric endocrine clinic of the second Hospital of Hebei Medical University from January 2015-07-07 to 2015-07-07. The concentrations of 25 (OH) D2X 25 (OH) D3, 25 (OH) D and 25 (OH) D in each serum were determined by LC-MS/MS, ECLIA and ELISA (IDS), respectively. SPSS17.0 software is used to compare the results of each method, linear correlation analysis and Bland-Altman consistency test. Results: 1 the data of serum 25 (OH) D measured by LC-MS/MS in 64 cases accord with normal distribution, the mean is 65.37 nmol / L, the standard deviation is 15.73 nmol / L; The results of ELISA and ECLIA were non-normal distribution, the median was 61.92 nmol / L ~ (60. 18) nmol/L.Wilcoxon test showed that there was no significant difference between the two methods. The average level of the three methods is close to that of the other three methods, the whole distribution of ELISA and LC-MS/MS is the same, and that of ECLIA is different from that of other two methods. 2 if the 25 (OH) D concentration of 50nmol / L (50-72.5nmol / L) is defined as vitamin D deficiency, the proportion of vitamin D deficiency in 64 children is 15.63% (ELISA),. 17.18% (LC-MS/MS), 31.25% (ECLIA); The rates of vitamin D deficiency were 57.82% (ELISA), 54.69% (LC-MS/MS), 39.06% (ECLIA). ELISA and LC-MS/MS results were the same, and the difference between ECLIA and LC-MS/MS was significant. 3 the linear regression equation and correlation coefficient between the two methods were: Y (ECLIA) = 1.32X (LC-MS/MS) -22.2 (r = 0.93); Y (ECLIA) = 1.05X (ELISA) -4.73 (r = 0.81); Y (ELISA) = 0.91X (LC-MS/MS) 5.79 (r = 0.84). There was a high positive correlation among the three methods. (4) Bland-Altman diagram of ECLIA and LC-MS/MS: the point of 4 / 64 is outside the 95% consistency limit. Compared with the two methods, the absolute value of the difference is 17.05 nmol / L, the ratio is the largest, the minimum is 1.3nmol / L, and the minimum is 1.3nmol / L, respectively. The point between ELISA and LC-MS/MS:4/64 lies outside the 95% consistency limit. The absolute value of the absolute value within the consistency limit is 18.17nmol / L, the ratio is the largest, the minimum is 1.3nmol / L, and the minimum is 1.3nmol / L, respectively. The point of ECLIA and ELISA:3/64 is outside the 95% consistency limit. The absolute value of the difference within the limit is 29.42 Nm / L, the ratio is the largest, the minimum is 1.5 卤0.6. Conclusion: when the total concentration of 25 (OH) D in serum is within a certain range and the content of 25 (OH) D2 is very low, there is a high positive correlation between ECLIA,LC-MS/MS,ELISA and serum 25 (OH) D, combined with box diagram and linear regression equation. Consistency analysis showed that the serum 25 (OH) D level measured by ECLIA was different from that of LC-MS/MS,ELISA to some extent, while LC-MS/MS,ELISA had little difference and good consistency.
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R446.6
[Abstract]:Objective: to compare the differences of simultaneous determination of 25 hydroxyvitamin D (25 (OH) D) in serum with three main methods: liquid chromatography-mass spectrometry (LC-MS/MS), electrochemiluminescence (ECLIA) and enzyme-linked immunosorbent assay (ELISA),). Consistency. Methods: 64 children with different levels of vitamin D were enrolled in the pediatric endocrine clinic of the second Hospital of Hebei Medical University from January 2015-07-07 to 2015-07-07. The concentrations of 25 (OH) D2X 25 (OH) D3, 25 (OH) D and 25 (OH) D in each serum were determined by LC-MS/MS, ECLIA and ELISA (IDS), respectively. SPSS17.0 software is used to compare the results of each method, linear correlation analysis and Bland-Altman consistency test. Results: 1 the data of serum 25 (OH) D measured by LC-MS/MS in 64 cases accord with normal distribution, the mean is 65.37 nmol / L, the standard deviation is 15.73 nmol / L; The results of ELISA and ECLIA were non-normal distribution, the median was 61.92 nmol / L ~ (60. 18) nmol/L.Wilcoxon test showed that there was no significant difference between the two methods. The average level of the three methods is close to that of the other three methods, the whole distribution of ELISA and LC-MS/MS is the same, and that of ECLIA is different from that of other two methods. 2 if the 25 (OH) D concentration of 50nmol / L (50-72.5nmol / L) is defined as vitamin D deficiency, the proportion of vitamin D deficiency in 64 children is 15.63% (ELISA),. 17.18% (LC-MS/MS), 31.25% (ECLIA); The rates of vitamin D deficiency were 57.82% (ELISA), 54.69% (LC-MS/MS), 39.06% (ECLIA). ELISA and LC-MS/MS results were the same, and the difference between ECLIA and LC-MS/MS was significant. 3 the linear regression equation and correlation coefficient between the two methods were: Y (ECLIA) = 1.32X (LC-MS/MS) -22.2 (r = 0.93); Y (ECLIA) = 1.05X (ELISA) -4.73 (r = 0.81); Y (ELISA) = 0.91X (LC-MS/MS) 5.79 (r = 0.84). There was a high positive correlation among the three methods. (4) Bland-Altman diagram of ECLIA and LC-MS/MS: the point of 4 / 64 is outside the 95% consistency limit. Compared with the two methods, the absolute value of the difference is 17.05 nmol / L, the ratio is the largest, the minimum is 1.3nmol / L, and the minimum is 1.3nmol / L, respectively. The point between ELISA and LC-MS/MS:4/64 lies outside the 95% consistency limit. The absolute value of the absolute value within the consistency limit is 18.17nmol / L, the ratio is the largest, the minimum is 1.3nmol / L, and the minimum is 1.3nmol / L, respectively. The point of ECLIA and ELISA:3/64 is outside the 95% consistency limit. The absolute value of the difference within the limit is 29.42 Nm / L, the ratio is the largest, the minimum is 1.5 卤0.6. Conclusion: when the total concentration of 25 (OH) D in serum is within a certain range and the content of 25 (OH) D2 is very low, there is a high positive correlation between ECLIA,LC-MS/MS,ELISA and serum 25 (OH) D, combined with box diagram and linear regression equation. Consistency analysis showed that the serum 25 (OH) D level measured by ECLIA was different from that of LC-MS/MS,ELISA to some extent, while LC-MS/MS,ELISA had little difference and good consistency.
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R446.6
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