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Immunological detection of thymine dimers in indigenous genomic DNA from pre-disinfection drinking water as an ultraviolet disinfection dosimeter
Environmental Science: Water Research & Technology ( IF 5 ) Pub Date : 2021-09-07 , DOI: 10.1039/d0ew00939c James Blyth 1 , Lucinda Hazell 1 , Michael R. Templeton 1
Environmental Science: Water Research & Technology ( IF 5 ) Pub Date : 2021-09-07 , DOI: 10.1039/d0ew00939c James Blyth 1 , Lucinda Hazell 1 , Michael R. Templeton 1
Affiliation
Culture-based methods are the primary methods used for the routine detection and enumeration of bacteria and viruses in water samples. In the context of ultraviolet (UV) disinfection, they are also the basis for reactor validation in drinking water treatment systems. However, the majority of microorganisms in pre-disinfection drinking water are not culturable. In UV disinfection, the DNA of both the culturable and non-culturable microbial populations will form pyrimidine dimers in response to UV photon absorbance. In this research an enzyme-linked immuno-sorbent assay (ELISA) was used to detect thymine dimers in the extractable genomic DNA (gDNA) from the total microbial population in pre-disinfection drinking water as a UV disinfection dosimeter. The method was first optimised using “naked” (extracted prior to UV exposure) and in vivo (extracted post UV exposure) E. coli gDNA, and then tested using water samples from UK drinking water treatment plants. Samples were exposed to up to 120 mJ cm−2 of monochromatic (253.7 nm) UV light using a collimated beam device and an ELISA was applied to the gDNA. This approach, once optimised, resulted in linear relationships between the assay response and UV dose. This shows that ELISA-based enumeration of thymine dimers in total extractable gDNA from a mixed species population has the potential to provide a direct, relatively quick, sampling-based means of monitoring the UV disinfection dose being delivered by operating UV disinfection systems in drinking water treatment plants, without the need to spike a biodosimeter into the water nor take reactors out of service. Molecular techniques measuring dimer formation may also offer the UV disinfection industry a method of demonstrating dose delivery where the culturing of target organisms is problematic.
中文翻译:
作为紫外线消毒剂量计,对来自预消毒饮用水的本地基因组 DNA 中胸腺嘧啶二聚体的免疫学检测
基于培养的方法是用于水样中细菌和病毒常规检测和计数的主要方法。在紫外线 (UV) 消毒的背景下,它们也是饮用水处理系统中反应器验证的基础。然而,消毒前饮用水中的大多数微生物是不可培养的。在紫外线消毒中,可培养和不可培养微生物种群的 DNA 将响应紫外线光子吸收形成嘧啶二聚体。在这项研究中,酶联免疫吸附测定 (ELISA) 被用于检测来自消毒前饮用水中总微生物群的可提取基因组 DNA (gDNA) 中的胸腺嘧啶二聚体,作为紫外线消毒剂量计。该方法首先使用“裸”(在紫外线照射前提取)和体内(紫外线暴露后提取)大肠杆菌gDNA,然后使用来自英国饮用水处理厂的水样进行测试。样品暴露于高达 120 mJ cm -2使用准直光束装置和 ELISA 将单色 (253.7 nm) 紫外光应用于 gDNA。这种方法经过优化后,会在检测响应和紫外线剂量之间产生线性关系。这表明,基于 ELISA 对来自混合物种种群的总可提取 gDNA 中胸腺嘧啶二聚体的计数有可能提供一种直接、相对快速、基于采样的方法来监测饮用水中操作紫外线消毒系统所提供的紫外线消毒剂量处理厂,无需将生物剂量计插入水中,也无需使反应器停止运行。测量二聚体形成的分子技术还可以为紫外线消毒行业提供一种证明剂量传递的方法,其中目标生物的培养存在问题。
更新日期:2021-09-08
中文翻译:
作为紫外线消毒剂量计,对来自预消毒饮用水的本地基因组 DNA 中胸腺嘧啶二聚体的免疫学检测
基于培养的方法是用于水样中细菌和病毒常规检测和计数的主要方法。在紫外线 (UV) 消毒的背景下,它们也是饮用水处理系统中反应器验证的基础。然而,消毒前饮用水中的大多数微生物是不可培养的。在紫外线消毒中,可培养和不可培养微生物种群的 DNA 将响应紫外线光子吸收形成嘧啶二聚体。在这项研究中,酶联免疫吸附测定 (ELISA) 被用于检测来自消毒前饮用水中总微生物群的可提取基因组 DNA (gDNA) 中的胸腺嘧啶二聚体,作为紫外线消毒剂量计。该方法首先使用“裸”(在紫外线照射前提取)和体内(紫外线暴露后提取)大肠杆菌gDNA,然后使用来自英国饮用水处理厂的水样进行测试。样品暴露于高达 120 mJ cm -2使用准直光束装置和 ELISA 将单色 (253.7 nm) 紫外光应用于 gDNA。这种方法经过优化后,会在检测响应和紫外线剂量之间产生线性关系。这表明,基于 ELISA 对来自混合物种种群的总可提取 gDNA 中胸腺嘧啶二聚体的计数有可能提供一种直接、相对快速、基于采样的方法来监测饮用水中操作紫外线消毒系统所提供的紫外线消毒剂量处理厂,无需将生物剂量计插入水中,也无需使反应器停止运行。测量二聚体形成的分子技术还可以为紫外线消毒行业提供一种证明剂量传递的方法,其中目标生物的培养存在问题。