Urea, the most abundant component in urine, cross-reacts with a commercial 8-OH-dG ELISA kit and contributes to overestimation of urinary 8-OH-dG
Urea, the most abundant component in urine, cross-reacts with a commercial 8-OH-dG ELISA kit and contributes to overestimation of urinary 8-OH-dG
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DOI：
10.1016/j.freeradbiomed.2009.02.017
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发表时间：
2009-07-01
影响因子：
7.4
通讯作者：
Ogawa, Yasutaka
中科院分区：
文献类型：
--
作者：
Song, Ming-Fen;Li, Yun-Shan;Ogawa, Yasutaka
尿8-OH-dG通常作为氧化应激的标志物进行分析。对于其分析，通常使用ELISA和HPLC方法，尽管经常讨论通过这些方法获得的数据的差异。为了澄清这个问题，我们通过反相HPLC分离人尿液，并通过ELISA方法测定每个馏分。除8-OH-dG级分外，在第一洗脱级分中观察到阳性反应。通过ELISA检测该级分中的组分。尿素被认为是在这一部分的责任成分。尿素在小鼠、大鼠和人类的尿液中以高浓度存在，其水平受许多因素影响。因此，需要进行某些改进，例如基于尿素含量的校正或折痕处理，以通过ELISA法准确分析尿8-OH-dG。此外，在4 ℃下进行的ELISA大大降低了对尿素的识别，并改善了8-OH-dG分析。(C)2009爱思唯尔公司All rights reserved.
Urinary 8-OH-dG is commonly analyzed as a marker of oxidative stress. For its analysis, ELISA and HPLC methods are generally used, although discrepancies in the data obtained by these methods have often been discussed. To clarify this problem, we fractionated human urine by reverse-phase HPLC and assayed each fraction by the ELISA method. In addition to the 8-OH-dG fraction, a positive reaction was observed in the first eluted fraction. The components in this fraction were examined by the ELISA. Urea was found to be the responsible component in this fraction. Urea is present in high concentrations in the urine of mice, rats, and humans, and its level is influenced by many factors. Therefore, certain improvements, such as a correction based on urea content or crease treatment, are required for the accurate analysis of urinary 8-OH-dG by the ELISA method. In addition, performance of the ELISA at 4 C reduced the recognition of urea considerably and improved the 8-OH-dG analysis. (C) 2009 Elsevier Inc. All rights reserved.