Improved single-swab sample preparation for recovering bacterial and phage DNA from human skin and wound microbiomes

Abstract Background Characterization of the skin and wound microbiome is of high biomedical interest, but is hampered by the low biomass of typical samples. While sample preparation from other microbiomes (e.g., gut) has been the subject of extensive optimization, procedures for skin and wound micro...

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Main Authors: Samuel Verbanic, Colin Y. Kim, John M. Deacon, Irene A. Chen
Format: Article
Language:English
Published: BMC 2019-09-01
Series:BMC Microbiology
Subjects:
Online Access:http://link.springer.com/article/10.1186/s12866-019-1586-4
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spelling doaj-7b74bf503dec4ff1b649a36170e861842020-11-25T03:25:58ZengBMCBMC Microbiology1471-21802019-09-0119111310.1186/s12866-019-1586-4Improved single-swab sample preparation for recovering bacterial and phage DNA from human skin and wound microbiomesSamuel Verbanic0Colin Y. Kim1John M. Deacon2Irene A. Chen3Department of Chemistry and Biochemistry, University of CaliforniaDepartment of Chemistry and Biochemistry, University of CaliforniaGoleta Valley Cottage Hospital, Ridley-Tree Center for Wound ManagementDepartment of Chemistry and Biochemistry, University of CaliforniaAbstract Background Characterization of the skin and wound microbiome is of high biomedical interest, but is hampered by the low biomass of typical samples. While sample preparation from other microbiomes (e.g., gut) has been the subject of extensive optimization, procedures for skin and wound microbiomes have received relatively little attention. Here we describe an improved method for obtaining both phage and microbial DNA from a single skin or wound swab, characterize the yield of DNA in model samples, and demonstrate the utility of this approach with samples collected from a wound clinic. Results We find a substantial improvement when processing wound samples in particular; while only one-quarter of wound samples processed by a traditional method yielded sufficient DNA for downstream analysis, all samples processed using the improved method yielded sufficient DNA. Moreover, for both skin and wound samples, community analysis and viral reads obtained through deep sequencing of clinical swab samples showed significant improvement with the use of the improved method. Conclusion Use of this method may increase the efficiency and data quality of microbiome studies from low-biomass samples.http://link.springer.com/article/10.1186/s12866-019-1586-4DNA purificationBacteriophageSkin microbiome
collection DOAJ
language English
format Article
sources DOAJ
author Samuel Verbanic
Colin Y. Kim
John M. Deacon
Irene A. Chen
spellingShingle Samuel Verbanic
Colin Y. Kim
John M. Deacon
Irene A. Chen
Improved single-swab sample preparation for recovering bacterial and phage DNA from human skin and wound microbiomes
BMC Microbiology
DNA purification
Bacteriophage
Skin microbiome
author_facet Samuel Verbanic
Colin Y. Kim
John M. Deacon
Irene A. Chen
author_sort Samuel Verbanic
title Improved single-swab sample preparation for recovering bacterial and phage DNA from human skin and wound microbiomes
title_short Improved single-swab sample preparation for recovering bacterial and phage DNA from human skin and wound microbiomes
title_full Improved single-swab sample preparation for recovering bacterial and phage DNA from human skin and wound microbiomes
title_fullStr Improved single-swab sample preparation for recovering bacterial and phage DNA from human skin and wound microbiomes
title_full_unstemmed Improved single-swab sample preparation for recovering bacterial and phage DNA from human skin and wound microbiomes
title_sort improved single-swab sample preparation for recovering bacterial and phage dna from human skin and wound microbiomes
publisher BMC
series BMC Microbiology
issn 1471-2180
publishDate 2019-09-01
description Abstract Background Characterization of the skin and wound microbiome is of high biomedical interest, but is hampered by the low biomass of typical samples. While sample preparation from other microbiomes (e.g., gut) has been the subject of extensive optimization, procedures for skin and wound microbiomes have received relatively little attention. Here we describe an improved method for obtaining both phage and microbial DNA from a single skin or wound swab, characterize the yield of DNA in model samples, and demonstrate the utility of this approach with samples collected from a wound clinic. Results We find a substantial improvement when processing wound samples in particular; while only one-quarter of wound samples processed by a traditional method yielded sufficient DNA for downstream analysis, all samples processed using the improved method yielded sufficient DNA. Moreover, for both skin and wound samples, community analysis and viral reads obtained through deep sequencing of clinical swab samples showed significant improvement with the use of the improved method. Conclusion Use of this method may increase the efficiency and data quality of microbiome studies from low-biomass samples.
topic DNA purification
Bacteriophage
Skin microbiome
url http://link.springer.com/article/10.1186/s12866-019-1586-4
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