Haugland Richard A, Siefring Shawn C, Wymer Larry J, Brenner Kristen P, Dufour Alfred P
US Environmental Protection Agency, National Exposure Research Laboratory, 26 W. Martin Luther King Dr., Cincinnati, OH 45268-1314, USA.
Water Res. 2005 Feb;39(4):559-68. doi: 10.1016/j.watres.2004.11.011. Epub 2004 Dec 24.
Cell densities of the fecal pollution indicator genus, Enterococcus, were determined by a rapid (3 h or less) quantitative polymerase chain reaction (QPCR) analysis method in 100 ml water samples collected from recreational beaches on Lake Michigan and Lake Erie during the summer of 2003. Measurements by this method were compared with counts of Enterococcus colony-forming units (CFU) determined by Method 1600 membrane filter (MF) analysis using mEI agar. The QPCR method had an estimated 95% confidence, minimum detection limit of 27 Enterococcus cells per sample in analyses of undiluted DNA extracts and quantitative analyses of multiple lake water samples, spiked with known numbers of these organisms, gave geometric mean results that were highly consistent with the spike levels. At both beaches, the geometric means of ambient Enterococcus concentrations in water samples, determined from multiple collection points during each sampling visit, showed approximately lognormal distributions over the study period using both QPCR and MF analyses. These geometric means ranged from 10 to 8548 cells by QPCR analysis and 1-2499 CFU by MF culture analysis in Lake Michigan (N=56) and from 8 to 8695 cells by QPCR and 3-1941 CFU by MF culture in Lake Erie (N=47). Regression analysis of these results showed a significant positive correlation between the two methods with an overall correlation coefficient (r) of 0.68.
2003年夏季,采用快速(3小时或更短时间)定量聚合酶链反应(QPCR)分析方法,对从密歇根湖和伊利湖休闲海滩采集的100毫升水样中粪便污染指示菌属肠球菌的细胞密度进行了测定。将该方法的测量结果与采用mEI琼脂的1600膜过滤(MF)分析法测定的肠球菌菌落形成单位(CFU)计数进行了比较。在未稀释DNA提取物的分析中,QPCR方法估计95%置信度下的最低检测限为每个样品27个肠球菌细胞,对添加已知数量这些微生物的多个湖水样品进行定量分析,得到的几何平均结果与添加水平高度一致。在两个海滩,每次采样时从多个采集点测定的水样中环境肠球菌浓度的几何平均值,在研究期间使用QPCR和MF分析均显示近似对数正态分布。在密歇根湖(N = 56),通过QPCR分析这些几何平均值范围为10至8548个细胞,通过MF培养分析为1至2499 CFU;在伊利湖(N = 47),通过QPCR为8至8695个细胞,通过MF培养为3至1941 CFU。对这些结果的回归分析表明,两种方法之间存在显著正相关,总体相关系数(r)为0.68。