Department of Microbiology, School of Biology, University of Barcelona, Barcelona, Spain.
J Appl Microbiol. 2010 Dec;109(6):1853-67. doi: 10.1111/j.1365-2672.2010.04830.x.
Molecular methods based on nucleic acid recognition and amplification are valuable tools to complement and support water management decisions. At present, these decisions are mostly supported by the principle of end-point monitoring for indicators and a small number of selected measured by traditional methods. Nucleic acid methods show enormous potential for identifying isolates from conventional culture methods, providing data on cultivable and noncultivable micro-organisms, informing on the presence of pathogens in waters, determining the causes of waterborne outbreaks, and, in some cases, detecting emerging pathogens. However, some features of water microbiology affect the performance of nucleic acid-based molecular techniques and thus challenge their suitability for routine water quality control. These features include the variable composition of target water samples, the generally low numbers of target micro-organisms, the variable water quality required for different uses and the physiological status or condition of such micro-organisms. The standardization of these molecular techniques is also an important challenge for its routine use in terms of accuracy (trueness and precision) and robustness (reproducibility and reliability during normal usage). Most of national and international water regulations recommend the application of standard methods, and any new technique must be validated respect to established methods and procedures. Moreover, molecular methods show a high cost-effectiveness value that limits its practicability on some microbial water analyses. However, new molecular techniques could contribute with new information or at least to supplement the limitation of traditional culture-based methods. Undoubtedly, challenges for these nucleic acid-based methods need to be identified and solved to improve their feasibility for routine microbial water monitoring.
基于核酸识别和扩增的分子方法是补充和支持水管理决策的有价值的工具。目前，这些决策主要依赖于终点监测指标的原则和少数传统方法测量的选定指标。核酸方法在识别常规培养方法中的分离株方面具有巨大的潜力，提供有关可培养和不可培养微生物的数据，提供水中病原体的存在信息，确定水源性暴发的原因，并在某些情况下检测新出现的病原体。然而，水微生物学的一些特征会影响基于核酸的分子技术的性能，从而挑战其在常规水质控制中的适用性。这些特征包括目标水样的组成变化、目标微生物的数量通常较低、不同用途所需的水质变化以及此类微生物的生理状态或条件。这些分子技术的标准化也是其常规使用在准确性（真实性和精密度）和稳健性（正常使用期间的可重复性和可靠性）方面的一个重要挑战。大多数国家和国际水法规都建议应用标准方法，任何新技术都必须针对既定方法和程序进行验证。此外，分子方法具有很高的成本效益价值，这限制了其在某些微生物水分析中的实用性。然而，新的分子技术可以提供新的信息，或者至少可以补充传统基于培养的方法的局限性。毫无疑问，这些基于核酸的方法需要确定和解决挑战，以提高其用于常规微生物水监测的可行性。
