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氧化状态的流行病学标志物:尿液2,3-二去甲-5,6-二氢-15-F2t-异前列腺素的酶联免疫吸附测定法与气相色谱/质谱分析法的比较

Epidemiological marker for oxidant status: comparison of the ELISA and the gas chromatography/mass spectrometry assay for urine 2,3-dinor-5,6-dihydro-15-F2t-isoprostane.

作者信息

Il'yasova Dora, Morrow Jason D, Ivanova Anastasia, Wagenknecht Lynne E

机构信息

Wake Forest University School of Medicine, Departments of Cancer Biology and Public Health Sciences, Winston-Salem, NC 27157, USA.

出版信息

Ann Epidemiol. 2004 Nov;14(10):793-7. doi: 10.1016/j.annepidem.2004.03.003.

Abstract

PURPOSE

A biomarker of oxidant status applicable to epidemiological research is essential to studying the relationship between free radicals and chronic disease risk. Gas chromatography with mass-spectrometry detection (GC/MS) is the gold standard for measurement of urinary F2-isoprostanes (F2-isoPs), a non-invasive marker of oxidant status. However, this method is laborious and costly, which prohibits its use in large epidemiological studies.

METHODS

We compared GC/MS assay with an inexpensive quick enzyme-linked immunoassay (ELISA) in measurements of 2,3-dinor-5,6-dihydro-15-F2t-isoprostane (F2-isoPM), an abundant beta-oxidation metabolite of 8-iso-prostaglandin-F2alpha. We measured F2-isoPM in urine of 52 participants of the Insulin Resistance Atherosclerosis Study by both methods.

RESULTS

The ELISA measurements showed approximately 30-fold greater mean and median (22.10, SD 12.92, and 18.49 ng/mg creatinine) than the GC/MS measurements (0.703, SD 0.468, and 0.597 ng/mg creatinine). We found low linear correlation (Pearson correlation coefficient 0.51; 95% CI, 0.28-0.70) and weak agreement in ranking subjects by tertiles (weighted Kappa statistic 0.34) between a GC/MS and ELISA.

CONCLUSIONS

We conclude that the current ELISA method is not a valid substitute for the GS/MS assay.

摘要

目的

一种适用于流行病学研究的氧化状态生物标志物对于研究自由基与慢性病风险之间的关系至关重要。气相色谱 - 质谱检测法(GC/MS)是测量尿F2 - 异前列腺素(F2 - isoPs)的金标准,F2 - isoPs是一种氧化状态的非侵入性标志物。然而,这种方法既费力又昂贵,这使其无法用于大型流行病学研究。

方法

我们在测量2,3 - 二去甲 - 5,6 - 二氢 - 15 - F2t - 异前列腺素(F2 - isoPM)时,将GC/MS检测法与一种廉价的快速酶联免疫吸附测定法(ELISA)进行了比较,F2 - isoPM是8 - 异前列腺素 - F2α的一种丰富的β - 氧化代谢产物。我们用这两种方法测量了胰岛素抵抗动脉粥样硬化研究中52名参与者尿液中的F2 - isoPM。

结果

ELISA测量结果显示,其平均值和中位数(分别为22.10,标准差12.92,以及18.49 ng/mg肌酐)比GC/MS测量结果(分别为0.703,标准差0.468,以及0.597 ng/mg肌酐)大约高30倍。我们发现GC/MS和ELISA之间的线性相关性较低(皮尔逊相关系数为0.51;95%置信区间为0.28 - 0.70),并且在按三分位数对受试者进行排名时一致性较弱(加权卡帕统计量为0.34)。

结论

我们得出结论,当前的ELISA方法不是GS/MS检测法的有效替代方法。

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