Introducing GUt Low-Density Array (GULDA) - a validated approach for qPCR-based intestinal microbial community analysis
Introducing GUt Low-Density Array (GULDA) - a validated approach for qPCR-based intestinal microbial community analysis
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DOI：
10.1111/1574-6968.12004
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发表时间：
2012-12-01
影响因子：
2.1
通讯作者：
Bahl, Martin I.
中科院分区：
文献类型：
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作者：
Bergstrom, Anders;Licht, Tine R.;Bahl, Martin I.
例如，由饮食、功能性食物、抗生素引起的人类肠道微生物群的改变，或作为年龄的函数发生的改变，现在已知与宿主健康相关。因此，强烈需要以快速和全面的方式检测这种改变的方法。在本研究中，我们开发并验证了一个高通量实时定量PCR为基础的分析平台，称为胃肠道低密度阵列（GULDA）。该平台设计用于同时分析在不同时间点从个体获得的粪便样品中31种不同微生物16 S rRNA基因靶标的丰度变化。靶基因代表肠道的五个主要细菌门（厚壁菌门、拟杆菌门、放线菌门、变形菌门和疣微菌门以及广古菌门）内的重要门、属、种或其他分类学组。为了证明GULDA的适用性，对从6名9和18个月大的健康婴儿获得的粪便样品进行分析，结果显示属于梭菌簇IV的细菌的相对丰度随时间显著增加（纤细梭菌组）和两歧双歧杆菌，同时丁酸梭菌丰度减少，肠杆菌科在9- 10年内有减少的趋势。月期间。
Alterations in the human gut microbiota caused, for example, by diet, functional foods, antibiotics, or occurring as a function of age are now known to be of relevance for host health. Therefore, there is a strong need for methods to detect such alterations in a rapid and comprehensive manner. In the present study, we developed and validated a high-throughput real-time quantitative PCR-based analysis platform, termed GUt Low-Density Array (GULDA). The platform was designed for simultaneous analysis of the change in the abundance of 31 different microbial 16S rRNA gene targets in fecal samples obtained from individuals at various points in time. The target genes represent important phyla, genera, species, or other taxonomic groups within the five predominant bacterial phyla of the gut, Firmicutes, Bacteroidetes, Actinobacteria, Proteobacteria, and Verrucomicrobia and also Euryarchaeota. To demonstrate the applicability of GULDA, analysis of fecal samples obtained from six healthy infants at both 9 and 18 months of age was performed and showed a significant increase over time of the relative abundance of bacteria belonging to Clostridial cluster IV (Clostridia leptum group) and Bifidobacterium bifidum and concurrent decrease in the abundance of Clostridium butyricum and a tendency for decrease in Enterobacteriaceae over the 9-month period.