Effects of sampling strategy and DNA extraction on human skin microbiome investigations.

Bjerre RD, Hugerth LW, Boulund F, Seifert M, Johansen JD, Engstrand L

Sci Rep 9 (1) 17287 [2019-11-21; online 2019-11-21]

The human skin is colonized by a wide array of microorganisms playing a role in skin disorders. Studying the skin microbiome provides unique obstacles such as low microbial biomass. The objective of this study was to establish methodology for skin microbiome analyses, focusing on sampling technique and DNA extraction. Skin swabs and scrapes were collected from 9 healthy adult subjects, and DNA extracted using 12 commercial kits. All 165 samples were sequenced using the 16S rRNA gene. Comparing the populations captured by eSwabs and scrapes, 99.3% of sequences overlapped. Using eSwabs yielded higher consistency. The success rate of library preparation applying different DNA extraction kits ranged from 39% to 100%. Some kits had higher Shannon alpha-diversity. Metagenomic shotgun analyses were performed on a subset of samples (N = 12). These data indicate that a reduction of human DNA from 90% to 57% is feasible without lowering the success of 16S rRNA library preparation and without introducing taxonomic bias. Using swabs is a reliable technique to investigate the skin microbiome. DNA extraction methodology is crucial for success of sequencing and adds a substantial amount of variation in microbiome analyses. Reduction of host DNA is recommended for interventional studies applying metagenomics.

DDLS Fellow

Luisa Hugerth

PubMed 31754146

DOI 10.1038/s41598-019-53599-z

Crossref 10.1038/s41598-019-53599-z

pmc: PMC6872721
pii: 10.1038/s41598-019-53599-z
figshare: 10.6084/m9.figshare.8319842.v1

Publications 9.5.0