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DOI | 10.1021/es401342b |
Filtration Recovery of Extracellular DNA from Environmental Water Samples | |
Liang, Zhanbei1; Keeley, Ann2 | |
发表日期 | 2013-08-20 |
ISSN | 0013-936X |
卷号 | 47期号:16页码:9324-9331 |
英文摘要 | qPCR methods are able to analyze DNA from microbes within hours of collecting water samples, providing the promptest notification and public awareness possible when unsafe pathogenic levels are reached. Health risk, however, may be overestimated by the presence of extracellular DNA (eDNA) that is corecovered by the filtration procedure which is the most commonly used method to concentrate target microbes from environmental waters. Using C. parvum 18S rRNA gene fragment as a representative of eDNA, we examined the impact of filters (types and pore sizes) and physiochemical properties of surface water samples on the recovery of spiked DNA. Our results indicated that binding affinities of various filter membranes were quantifiably different for eDNA fragments with the polycarbonate (PC) binding the least and mixed cellulose acetate and cellulose nitrate (MCE) binding the most as evidenced by up to 16% recovery of the spiked plasmid DNA with a pore size of 0.2 mu m. Water quality parameters also had a distinct influence on the recovery of eDNA which was enhanced by the presence of high total suspended solid (TSS) concentrations and reduced pH. At pH 53, with 150 mg/L of clay, DNA recovery was increased to as much as 18%. By shielding the negative charge, thus increasing the interaction of DNA and colloids, the increase of Na+ and Ca2+ concentrations resulted in more DNA binding and consequently more recovery from environmental water samples. Therefore, in addition to analytical uncertainties, potential differences in qPCR data from filtered waters characterized with low pH and high TSS and ionic strength should be considered in pollution assessments. |
语种 | 英语 |
WOS记录号 | WOS:000323471700041 |
来源期刊 | ENVIRONMENTAL SCIENCE & TECHNOLOGY
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来源机构 | 美国环保署 |
文献类型 | 期刊论文 |
条目标识符 | http://gcip.llas.ac.cn/handle/2XKMVOVA/57186 |
作者单位 | 1.US EPA, Natl Res Council, Ada, OK 74820 USA; 2.US EPA, Natl Risk Management Res Lab, Ada, OK 74820 USA |
推荐引用方式 GB/T 7714 | Liang, Zhanbei,Keeley, Ann. Filtration Recovery of Extracellular DNA from Environmental Water Samples[J]. 美国环保署,2013,47(16):9324-9331. |
APA | Liang, Zhanbei,&Keeley, Ann.(2013).Filtration Recovery of Extracellular DNA from Environmental Water Samples.ENVIRONMENTAL SCIENCE & TECHNOLOGY,47(16),9324-9331. |
MLA | Liang, Zhanbei,et al."Filtration Recovery of Extracellular DNA from Environmental Water Samples".ENVIRONMENTAL SCIENCE & TECHNOLOGY 47.16(2013):9324-9331. |
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