Fromin N, Hamelin J, Tarnawski S, Roesti D, Jourdain-Miserez K, Forestier N, Teyssier-Cuvelle S, Gillet F, Aragno M, Rossi P
Microbiology Laboratory, University of Neuchâtel, Switzerland.
Environ Microbiol. 2002 Nov;4(11):634-43. doi: 10.1046/j.1462-2920.2002.00358.x.
Technical developments in molecular biology have found extensive applications in the field of microbial ecology. Among these techniques, fingerprinting methods such as denaturing gel electrophoresis (DGE, including the three options: DGGE, TGGE and TTGE) has been applied to environmental samples over this last decade. Microbial ecologists took advantage of this technique, originally developed for the detection of single mutations, for the analysis of whole bacterial communities. However, until recently, the results of these high quality fingerprinting patterns were restricted to a visual interpretation, neglecting the analytical potential of the method in terms of statistical significance and ecological interpretation. A brief recall is presented here about the principles and limitations of DGE fingerprinting analysis, with an emphasis on the need of standardization of the whole analytical process. The main content focuses on statistical strategies for analysing the gel patterns, from single band examination to the analysis of whole fingerprinting profiles. Applying statistical method make the DGE fingerprinting technique a promising tool. Numerous samples can be analysed simultaneously, permitting the monitoring of microbial communities or simply bacterial groups for which occurrence and relative frequency are affected by any environmental parameter. As previously applied in the fields of plant and animal ecology, the use of statistics provides a significant advantage for the non-ambiguous interpretation of the spatial and temporal functioning of microbial communities.
分子生物学的技术发展在微生物生态学领域得到了广泛应用。在这些技术中,指纹图谱方法,如变性凝胶电泳(DGE,包括DGGE、TGGE和TTGE三种选择)在过去十年中已应用于环境样品。微生物生态学家利用这种最初为检测单基因突变而开发的技术来分析整个细菌群落。然而,直到最近,这些高质量指纹图谱的结果仅限于视觉解读,忽略了该方法在统计显著性和生态学解释方面的分析潜力。本文简要回顾了DGE指纹图谱分析的原理和局限性,重点强调了整个分析过程标准化的必要性。主要内容聚焦于分析凝胶图谱的统计策略,从单带检测到整个指纹图谱的分析。应用统计方法使DGE指纹图谱技术成为一种有前景的工具。可以同时分析大量样品,从而能够监测微生物群落或仅仅是那些其出现和相对频率受任何环境参数影响的细菌类群。正如之前在植物和动物生态学领域的应用一样,统计学的使用为明确解读微生物群落的时空功能提供了显著优势。