Galluzzi Luca, Magnani Mauro, Saunders Nick, Harms Carsten, Bruce Ian James
Istituto di Chimica Biologica G. Fornaini, Università degli Studi Carlo Bo, Via Saffi, 2, 61029 Urbino, Italy.
Sci Prog. 2007;90(Pt 1):29-50. doi: 10.3184/003685007780440521.
Traditionally the detection of microbial pathogens in clinical, environmental or food samples has commonly needed the prelevation of cells by culture before the application ofthe detection strategy. This is done to increase cell number thereby overcoming problems associated with the sensitivity of classical detection strategies. However, culture-based methods have the disadvantages of taking longer, usually are more complex and require skilled personnel as well as not being able to detect viable but non cultivable microbial species. A number of molecular methods have been developed in the last 10 to 15 years to overcome these issues and to facilitate the rapid, accurate, sensitive and cost effective identification and enumeration of microorganisms which are designed to replace and/or support classical approaches to microbial detection. Amongst these new methods, ones based on the polymerase chain reaction and nucleic acid hybridization have been shown to be particularly suitable for this purpose. This review generally summarizes some of the current and emerging nucleic acid based molecular approaches for the detection, discrimination andquantification ofmicrobes in environmental, food and clinical samples and includes reference to the recently developing areas of microfluidics and nanotechnology "Lab-on-a-chip".
传统上,在临床、环境或食品样本中检测微生物病原体时,通常需要在应用检测策略之前通过培养来富集细胞。这样做是为了增加细胞数量,从而克服与传统检测策略灵敏度相关的问题。然而,基于培养的方法存在耗时较长、通常更为复杂且需要专业人员等缺点,同时还无法检测活的但不可培养的微生物种类。在过去的10到15年里,人们开发了许多分子方法来克服这些问题,并促进对微生物进行快速、准确、灵敏且经济高效的鉴定和计数,这些方法旨在替代和/或支持传统的微生物检测方法。在这些新方法中,基于聚合酶链反应和核酸杂交的方法已被证明特别适用于此目的。本综述总体上总结了一些当前以及新兴的基于核酸的分子方法,用于检测、区分和定量环境、食品和临床样本中的微生物,并且提及了微流控和纳米技术“芯片实验室”等最近的发展领域。