Comparison of four DNA extraction methods for 16s rRNA microbiota profiling of human faecal samples

四种 DNA 提取方法对人类粪便样本 16s rRNA 微生物群分析的比较

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作者:James Sinclair #, Nicholas P West, Amanda J Cox #

Objective

Growth in large population-based studies assessing contributions of the gut microbiota to health and disease requires high-throughput sample processing and analysis

Results

Diversity metrics did not differ between extraction methods. Differences in the relative abundance of key phyla, including a significantly lower abundance of the Firmicutes (p = 0.004) and higher relative abundance of the Bacteroidetes (p = 0.005) and Proteobacteria (p = 0.008) phyla were noted where the DNA extraction did not include additional chemical and mechanical lysis. Principal coordinate analysis of family and genera level data also suggested a potential for sample pre-processing to impact microbial composition. Observations of the potential for skewed microbial composition profiles from samples prepared using a semi-automated DNA extraction kit without additional sample pre-processing highlights a need for consideration of standardisation of methodological approaches to increase the comparability of microbial compositional data.

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