Bissonnette Luc, Maheux Andrée F, Bergeron Michel G
Centre de recherche en infectiologie de l'Université Laval, Axe maladies infectieuses et immunitaires, Centre de recherche du CHU de Québec-Université Laval, Québec City, QC, Canada, G1V 4G2.
AFM Water Consulting, Québec City, QC, Canada.
Methods Mol Biol. 2017;1620:141-151. doi: 10.1007/978-1-4939-7060-5_9.
The microbial assessment of potable/drinking water is done to ensure that the resource is free of fecal contamination indicators or waterborne pathogens. Culture-based methods for verifying the microbial safety are limited in the sense that a standard volume of water is generally tested for only one indicator (family) or pathogen.In this work, we describe a membrane filtration-based molecular microbiology method, CRENAME (Concentration Recovery Extraction of Nucleic Acids and Molecular Enrichment), exploiting molecular enrichment by whole genome amplification (WGA) to yield, in less than 4 h, a nucleic acid preparation which can be repetitively tested by real-time PCR for example, to provide multiparametric presence/absence tests (1 colony forming unit or microbial particle per standard volume of 100-1000 mL) for bacterial or protozoan parasite cells or particles susceptible to contaminate potable/drinking water.
对饮用水进行微生物评估是为了确保该水源不含粪便污染指标或水源性病原体。基于培养的微生物安全验证方法存在局限性,因为通常仅针对一种指标(类别)或病原体对标准体积的水进行检测。在本研究中,我们描述了一种基于膜过滤的分子微生物学方法,即CRENAME(核酸浓度回收提取与分子富集法),该方法利用全基因组扩增(WGA)进行分子富集,在不到4小时的时间内获得核酸制剂,例如可通过实时PCR对其进行重复检测,以针对可能污染饮用水的细菌或原生动物寄生虫细胞或颗粒提供多参数存在/不存在检测(每100 - 1000 mL标准体积中有1个菌落形成单位或微生物颗粒)。