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城市饮用水中DNA浓度的荧光测定法。

Fluorometric determination of the DNA concentration in municipal drinking water.

作者信息

McCoy W F, Olson B H

出版信息

Appl Environ Microbiol. 1985 Apr;49(4):811-7. doi: 10.1128/aem.49.4.811-817.1985.

Abstract

DNA concentrations in municipal drinking water samples were measured by fluorometry, using Hoechst 33258 fluorochrome. The concentration, extraction, and detection methods used were adapted from existing techniques. The method is reproducible, fast, accurate, and simple. The amounts of DNA per cell for five different bacterial isolates obtained from drinking water samples were determined by measuring DNA concentration and total cell concentration (acridine orange epifluorescence direct cell counting) in stationary pure cultures. The relationship between DNA concentration and epifluorescence total direct cell concentration in 11 different drinking water samples was linear and positive; the amounts of DNA per cell in these samples did not differ significantly from the amounts in pure culture isolates. We found significant linear correlations between DNA concentration and colony-forming unit concentration, as well as between epifluorescence direct cell counts and colony-forming unit concentration. DNA concentration measurements of municipal drinking water samples appear to monitor changes in bacteriological quality at least as well as total heterotrophic plate counting and epifluorescence direct cell counting.

摘要

使用Hoechst 33258荧光染料通过荧光测定法测量市政饮用水样本中的DNA浓度。所使用的浓度、提取和检测方法均改编自现有技术。该方法具有可重复性、快速、准确且简单的特点。通过测量静止纯培养物中的DNA浓度和总细胞浓度(吖啶橙落射荧光直接细胞计数),确定了从饮用水样本中获得的五种不同细菌分离株的每细胞DNA量。11个不同饮用水样本中DNA浓度与落射荧光总直接细胞浓度之间的关系呈线性且为正相关;这些样本中的每细胞DNA量与纯培养分离株中的量没有显著差异。我们发现DNA浓度与菌落形成单位浓度之间以及落射荧光直接细胞计数与菌落形成单位浓度之间存在显著的线性相关性。市政饮用水样本的DNA浓度测量似乎至少与总异养平板计数和落射荧光直接细胞计数一样,能够监测细菌学质量的变化。

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