Molecular biological methods for studying the gut microbiota:: the EU human gut flora project
Molecular biological methods for studying the gut microbiota:: the EU human gut flora project
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DOI：
10.1079/bjn/2002539
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发表时间：
2002-05-01
影响因子：
3.6
通讯作者：
de Vos, W
中科院分区：
文献类型：
--
作者：
Blaut, M;Collins, MD;de Vos, W
七个欧洲实验室在一个联合项目（FAIR CT 97 -3035）中合作，开发、改进和应用分子方法，以促进阐明人类肠道微植物群的复杂组成，并设计监测肠道植物群对饮食反应的可靠方法。一个广泛的数据库，用于跟踪肠道细菌的16 S rRNA序列的测序新的粪便分离株的16 S rRNA基因和克隆的粪便DNA的聚合酶链反应（PCR）扩增来自不同年龄组的受试者。分析表明，人类肠道中存在的不同物种（多样性）的数量随着年龄的增长而增加。所产生的序列信息为设计16 S rRNA指导的寡核苷酸探针提供了基础，以特异性地检测各种系统发育层次的细菌。测试探针的特异性，并用于全细胞和斑点杂交。在几项研究中证明了所开发的方法的适用性，并描述了主要结果。
Seven European laboratories co-operated in a joint project (FAIR CT97-3035) to develop, refine and apply molecular methods towards facilitating elucidation of the complex composition of the human intestinal microflora and to devise robust methodologies for monitoring the gut flora in response to diet. An extensive database of 16S rRNA sequences for tracking intestinal bacteria was generated by sequencing the 16S rRNA genes of new faecal isolates and of clones obtained by amplification with polymerase chain reaction (PCR) on faecal DNA from subjects belonging to different age groups. The analyses indicated that the number of different species (diversity) present in the human gut increased with age. The sequence information generated, provided the basis for design of 16S rRNA-directed oligonucleotide probes to specifically detect bacteria at various levels of phylogenetic hierarchy. The probes were tested for their specificity and used in whole-cell and dot-blot hybridisations. The applicability of the developed methods was demonstrated in several studies and the major outcomes are described.