Andersson Anders F, Lindberg Mathilda, Jakobsson Hedvig, Bäckhed Fredrik, Nyrén Pål, Engstrand Lars
Swedish Institute for Infectious Disease Control, Solna, Sweden.
PLoS One. 2008 Jul 30;3(7):e2836. doi: 10.1371/journal.pone.0002836.
Humans host complex microbial communities believed to contribute to health maintenance and, when in imbalance, to the development of diseases. Determining the microbial composition in patients and healthy controls may thus provide novel therapeutic targets. For this purpose, high-throughput, cost-effective methods for microbiota characterization are needed. We have employed 454-pyrosequencing of a hyper-variable region of the 16S rRNA gene in combination with sample-specific barcode sequences which enables parallel in-depth analysis of hundreds of samples with limited sample processing. In silico modeling demonstrated that the method correctly describes microbial communities down to phylotypes below the genus level. Here we applied the technique to analyze microbial communities in throat, stomach and fecal samples. Our results demonstrate the applicability of barcoded pyrosequencing as a high-throughput method for comparative microbial ecology.
人类体内寄生着复杂的微生物群落，据信这些群落有助于维持健康，而当群落失衡时，则会导致疾病的发生。因此，确定患者和健康对照者体内的微生物组成可能会提供新的治疗靶点。为此，需要高通量、具有成本效益的微生物群特征分析方法。我们采用了对16S rRNA基因高变区进行454焦磷酸测序，并结合样本特异性条形码序列，这使得我们能够在有限的样本处理条件下对数百个样本进行平行深入分析。计算机模拟表明，该方法能够准确描述低至属以下系统发育型的微生物群落。在这里，我们应用该技术分析了咽喉、胃部和粪便样本中的微生物群落。我们的结果证明了条形码焦磷酸测序作为一种高通量方法在比较微生物生态学中的适用性。
