Molecular monitoring of succession of bacterial communities in human neonates
Molecular monitoring of succession of bacterial communities in human neonates
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DOI：
10.1128/aem.68.1.219-226.2002
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发表时间：
2002-01-01
影响因子：
4.4
通讯作者：
Akkermans, ADL
中科院分区：
文献类型：
--
作者：
Favier, CF;Vaughan, EE;Akkermans, ADL
通过PCR和变性梯度凝胶电泳（DGGE）监测粪便样本中的16 S核糖体DNA（rDNA）多样性，并通过分析主要核糖体类型的序列，在两个健康婴儿的细菌群落的建立超过生命的前10个月进行了检查。通过分析每日或每周的粪便样品，获得婴儿肠道中优势种群的DGGE图谱。分娩后，无菌婴儿胃肠道迅速定植，并监测每个生态系统中的细菌演替。在生命的最初几天，这些特征很简单，但随着两个婴儿的细菌多样性随着时间的推移而增加，它们变得更加复杂。从婴儿粪便中扩增的16 S rDNA片段的克隆文库的构建，这些文库允许通过比较DNA序列分析鉴定细菌类型;鉴定的细菌包括双歧杆菌属，瘤胃球菌属，肠球菌属，梭菌属和肠杆菌属的成员。与双歧杆菌属和瘤胃球菌属最密切相关的物种特别是基于随时间的稳定性和数量（如通过条带的强度估计的）主导肠道微生物群。然而，34个克隆的rDNA序列中有19个与已知细菌或数据库中克隆的序列的同源性低于97%。这项研究表明，使用PCR-DGGE和16 S rDNA序列分析一起导致在婴儿肠道生态系统中的细菌定植的动态描述，并允许可视化的细菌，难以培养或通过其他方法检测。
The establishment of bacterial communities in two healthy babies was examined for more than the first 10 months of life by monitoring 16S ribosomal DNA (rDNA) diversity in fecal samples by PCR and denaturing gradient gel electrophoresis (DGGE) and by analyzing the sequences of the major ribotypes. DGGE profiles of the dominant populations in the intestines of the infants were obtained by analyzing daily or weekly fecal samples. After delivery, the germfree infant gastrointestinal tracts were rapidly colonized, and the succession of bacteria in each ecosystem was monitored. During the first few days of life the profiles were simple, but they became more complex as the bacterial diversity increased with time in both babies. Clone libraries of amplified 16S rDNA fragments from baby feces were constructed, and these libraries allowed identification of the bacterial types by comparative DNA sequence analysis; the bacteria identified included members of the genera Bifidobacterium, Ruminococcus, Enterococcus, Clostridium, and Enterobacter. Species most closely related to the genera Bifidobacterium and Ruminococcus in particular dominated the intestinal microbiota based on the stability over time and the numbers, as estimated by the intensities of the bands. However, 19 of the 34 cloned rDNA sequences exhibited less than 97% identity with sequences of known bacteria or cloned sequences in databases. This study showed that using PCR-DGGE and 16S rDNA sequence analysis together resulted in a dynamic description of bacterial colonization in the infant intestinal ecosystem and allowed visualization of bacteria that are difficult to cultivate or to detect by other methods.