期刊
JOURNAL OF MICROBIOLOGICAL METHODS
卷 119, 期 -, 页码 239-242出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.mimet.2015.11.001
关键词
Metagenomics; Microbiota; Microbiome; 16s sequencing; Prokaryotic DNA; Eukatyotic human DNA; Separation; Purification
资金
- Broad Medical Research Program (BMRP-CCFA) [335929 IBD-0381]
- State of Montana, Montana Department of Commerce Biomedical Research Grant [14-51-BMR-002]
- St. Vincent Healthcare
- Montana State University-Billings
Microbial metagenomics are hindered in clinical tissue samples as a result of the large relative amount of human DNA in relation to microbial DNA acting as competitive inhibitors of downstream applications. We evaluated the LOOXSTER (R) Enrichment Kit to separate eukaryotic and prokaryotic DNA in submucosal intestinal tissue samples having a low microbial biomass and to determine the effects of enrichment on 16s rRNA microbiota sequencing. The enrichment kit reduced the amount of human DNA in the samples 40-70% resulting in a 3.5-fold increase in the number of 16s bacterial gene sequences detected on the Illumina MiSeq platform. This increase was accompanied by the detection of 41 additional bacterial genera and 94 tentative species. The additional bacterial taxa detected accounted for as much as 25% of the total bacterial population that significantly altered the relative prevalence and composition of the intestinal microbiota. The ability to reduce the competitive inhibition created by human DNA and the concentration of bacterial DNA may allow metagenomics to be performed on complex tissues containing a low bacterial biomass. (C) 2015 Elsevier B.V. All rights reserved.
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