Huys Geert, Vanhoutte Tom, Vandamme Peter
Laboratory of Microbiology, Faculty of Sciences, Ghent University, K.L. Ledeganckstraat 35, 9000 Ghent, Belgium.
Interdiscip Perspect Infect Dis. 2008;2008:597603. doi: 10.1155/2008/597603. Epub 2008 Dec 14.
Sequence-dependent electrophoresis (SDE) fingerprinting techniques such as denaturing gradient gel electrophoresis (DGGE) have become commonplace in the field of molecular microbial ecology. The success of the SDE technology lays in the fact that it allows visualization of the predominant members of complex microbial ecosystems independent of their culturability and without prior knowledge on the complexity and diversity of the ecosystem. Mainly using the prokaryotic 16S rRNA gene as PCR amplification target, SDE-based community fingerprinting turned into one of the leading molecular tools to unravel the diversity and population dynamics of human intestinal microbiota. The first part of this review covers the methodological concept of SDE fingerprinting and the technical hurdles for analyzing intestinal samples. Subsequently, the current state-of-the-art of DGGE and related techniques to analyze human intestinal microbiota from healthy individuals and from patients with intestinal disorders is surveyed. In addition, the applicability of SDE analysis to monitor intestinal population changes upon nutritional or therapeutic interventions is critically evaluated.
诸如变性梯度凝胶电泳(DGGE)之类的序列依赖性电泳(SDE)指纹技术在分子微生物生态学领域已变得很常见。SDE技术的成功在于它能够可视化复杂微生物生态系统中的主要成员,而无需考虑它们的可培养性,也无需事先了解生态系统的复杂性和多样性。主要以原核生物16S rRNA基因作为PCR扩增靶点,基于SDE的群落指纹分析成为揭示人类肠道微生物群多样性和种群动态的主要分子工具之一。本综述的第一部分涵盖了SDE指纹分析的方法概念以及分析肠道样本的技术障碍。随后,调查了用于分析健康个体和肠道疾病患者的人类肠道微生物群的DGGE及相关技术的当前技术水平。此外,还对SDE分析在监测营养或治疗干预后肠道菌群变化方面的适用性进行了批判性评估。