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饮用水中超低水平DNA变化的检测:具有流行病学重要意义的发现。

Detection of ultra-low levels of DNA changes by drinking water: epidemiologically important finding.

作者信息

Kumari Parmila, Kamiseki Meiko, Biyani Manish, Suzuki Miho, Nemoto Naoto, Aita Takuyo, Nishigaki Koichi

机构信息

Graduate School of Science and Engineering, Saitama University, 255 Shimo-Okubo, Sakura-Ku, Saitama-City, Saitama 338-8570, Japan and Department of Bioengineering, University of Tokyo 2-11-16, Yayoi, Bunkyo-Ku, Tokyo 113-8656, Japan

Graduate School of Science and Engineering, Saitama University, 255 Shimo-Okubo, Sakura-Ku, Saitama-City, Saitama 338-8570, Japan and Department of Bioengineering, University of Tokyo 2-11-16, Yayoi, Bunkyo-Ku, Tokyo 113-8656, Japan.

出版信息

J Biochem. 2015 Feb;157(2):113-9. doi: 10.1093/jb/mvu072. Epub 2014 Nov 17.

Abstract

The safety of drinking water is essential to our health. In this context, the mutagenicity of water needs to be checked strictly. However, from the methodological limit, the lower concentration (less than parts per million) of mutagenicity could not be detected, though there have been of interest in the effect of less concentration mutagens. Here, we describe a highly sensitive mutation assay that detects mutagens at the ppb level, termed genome profiling-based mutation assay (GPMA). This consists of two steps; (i) Escherichia coli culture in the medium with/without mutagens and (ii) Genome profiling (GP) method (an integrated method of random PCR, temperature gradient gel electrophoresis and computer-aided normalization). Owing to high sensitivity of this method, very low concentration of mutagens in tap water could be directly detected without introducing burdensome concentration processes, enabling rapid measurement of low concentration samples. Less expectedly, all of the tap waters tested (22 samples) were shown to be significantly mutagenic while mineral waters were not. Resultantly, this article informs two facts that the GPMA method is competent to measure the mutagenicity of waters directly and the experimental results supported the former reports that the city tap waters contain very low level of mutagenicity reagent trihalomethanes.

摘要

饮用水安全对我们的健康至关重要。在此背景下,水的致突变性需要严格检测。然而,由于方法学的限制,尽管低浓度(低于百万分之一)的致突变性备受关注,但仍无法检测到。在此,我们描述了一种高灵敏度的突变检测方法,可在十亿分之一水平检测致突变物,称为基于基因组图谱的突变检测法(GPMA)。该方法包括两个步骤:(i)在含有/不含有致突变物的培养基中培养大肠杆菌,以及(ii)基因组图谱(GP)方法(一种随机PCR、温度梯度凝胶电泳和计算机辅助归一化的综合方法)。由于该方法灵敏度高,无需进行繁琐的浓缩过程即可直接检测自来水中极低浓度的致突变物,从而能够快速测量低浓度样品。出乎意料的是,所有测试的自来水(22个样本)均显示出显著的致突变性,而矿泉水则没有。结果表明,本文揭示了两个事实:GPMA方法能够直接测量水的致突变性,实验结果支持了之前关于城市自来水含有极低水平致突变试剂三卤甲烷的报道。

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