Validating T-RFLP as a sensitive and high-throughput approach to assess bacterial diversity patterns in human anterior nares
Amélia Camarinha-Silva
Microbial Interactions and Processes Research Group, Department of Medical Microbiology, Helmholtz Centre for Infection Research, Braunschweig, Germany
Search for more papers by this authorMelissa L. Wos-Oxley
Microbial Interactions and Processes Research Group, Department of Medical Microbiology, Helmholtz Centre for Infection Research, Braunschweig, Germany
Search for more papers by this authorRuy Jáuregui
Microbial Interactions and Processes Research Group, Department of Medical Microbiology, Helmholtz Centre for Infection Research, Braunschweig, Germany
Search for more papers by this authorKarsten Becker
Institute of Medical Microbiology, University Hospital Münster, Münster, Germany
Search for more papers by this authorCorresponding Author
Dietmar H. Pieper
Microbial Interactions and Processes Research Group, Department of Medical Microbiology, Helmholtz Centre for Infection Research, Braunschweig, Germany
Correspondence: Dietmar H. Pieper, Microbial Interactions and Processes Research Group, Department of Medical Microbiology, Helmholtz Centre for Infection Research, Inhoffenstr. 7, 38124 Braunschweig, Germany. Tel.: +49 531 6181 4200; fax: +49 531 6181 4499; e-mail: [email protected]Search for more papers by this authorAmélia Camarinha-Silva
Microbial Interactions and Processes Research Group, Department of Medical Microbiology, Helmholtz Centre for Infection Research, Braunschweig, Germany
Search for more papers by this authorMelissa L. Wos-Oxley
Microbial Interactions and Processes Research Group, Department of Medical Microbiology, Helmholtz Centre for Infection Research, Braunschweig, Germany
Search for more papers by this authorRuy Jáuregui
Microbial Interactions and Processes Research Group, Department of Medical Microbiology, Helmholtz Centre for Infection Research, Braunschweig, Germany
Search for more papers by this authorKarsten Becker
Institute of Medical Microbiology, University Hospital Münster, Münster, Germany
Search for more papers by this authorCorresponding Author
Dietmar H. Pieper
Microbial Interactions and Processes Research Group, Department of Medical Microbiology, Helmholtz Centre for Infection Research, Braunschweig, Germany
Correspondence: Dietmar H. Pieper, Microbial Interactions and Processes Research Group, Department of Medical Microbiology, Helmholtz Centre for Infection Research, Inhoffenstr. 7, 38124 Braunschweig, Germany. Tel.: +49 531 6181 4200; fax: +49 531 6181 4499; e-mail: [email protected]Search for more papers by this authorAbstract
While recent works aimed to thoroughly characterize the bacterial community of the human anterior nares of a few candidates, this work sought to analyse a greater cross-section by sampling 100 volunteers. After optimizing and validating the method of terminal restriction fragment length polymorphism against six previously pyrosequenced samples, abundant species could be discriminated and their relative abundances measured in a high-throughput manner. The 100 volunteers could be statistically clustered into 12 groups, where two-thirds of volunteers shared more than 40% similarity in respect to their bacterial community structure, while the remaining third clustered into smaller groups being dominated by Dolosigranulum pigrum, Moraxella spp. or Staphylococcus aureus. Moraxella spp. was present predominantly in women rather than in men. The use of network analysis charting bacterial ecological co-occurrences revealed new evidence of likely positive associations between some core human nasal species. So, in the age of post ‘omics’ and ‘deep sequencing’, there is still a place for these well-tried and well-tested methods that can offer a rapid, reproducible and economical alternative, whereby also yielding valuable new information.
Supporting Information
Filename | Description |
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fem1197-sup-0001-FigureS1.tifimage/tif, 77.1 KB | Fig. S1. Non-metric multidimensional scaling plots illustrating similarities in the global bacterial community structure of the anterior nares of six volunteers as analyzed by T-RFLP (denoted as T) compared to 454-pyrosequencing data (denoted as P) (volunteers 3, 10, 13, 15, 17, 18 as described by Wos-Oxley et al., 2010). |
fem1197-sup-0002-FigureS2.tifimage/tif, 319 KB | Fig. S2. Comparison of the % abundance of species-of-interest in the global bacterial community structure of the anterior nares of six volunteers as analyzed by T-RFLP (black) compared to 454-pyrosequencing data (grey) (volunteers 3, 10, 13, 15, 17, 18 as described by Wos-Oxley et al., 2010). |
fem1197-sup-0003-TableS1.docWord document, 35.5 KB | Table S1. Primers used in this study. |
fem1197-sup-0004-TableS2.docWord document, 94 KB | Table S2. Characteristics (gender, age, smoking habits, ownership of pets) of each of the 100 volunteers used in this study. |
fem1197-sup-0005-TableS3.docWord document, 49.5 KB | Table S3. Observed terminal restriction fragments (in bp) after sequential digestion with AseI, TspRI and ApekI in samples collected from 100 volunteers. |
fem1197-sup-0006-TableS4.docWord document, 44.5 KB | Table S4. Limit of detection and quantification of S. aureus (in those volunteers that are S. aureus carriers) using RTQ-PCR. |
fem1197-sup-0007-TableS5.docWord document, 40 KB | Table S5. Summary of those groups determined by SIMPROF and their characteristic profiles. |
Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.
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