Kwan, K. and Cooper, M. and La Duc, M. T. and Vaishampayan, P. and Stam, C. and Benardini, J. N. and Scalzi, G. and Moissl-Eichinger, C. and Venkateswaran, K. (2011) Evaluation of Procedures for the Collection, Processing, and Analysis of Biomolecules from Low-Biomass Surfaces. APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 77 (9). pp. 2943-2953. ISSN 0099-2240,
Full text not available from this repository. (Request a copy)Abstract
To comprehensively assess microbial diversity and abundance via molecular-analysis-based methods, procedures for sample collection, processing, and analysis were evaluated in depth. A model microbial community (MMC) of known composition, representative of a typical low-biomass surface sample, was used to examine the effects of variables in sampling matrices, target cell density/molecule concentration, and cryogenic storage on the overall efficacy of the sampling regimen. The MMC used in this study comprised 11 distinct species of bacterial, archaeal, and fungal lineages associated with either spacecraft or clean-room surfaces. A known cellular density of MMC was deposited onto stainless steel coupons, and after drying, a variety of sampling devices were used to recover cells and biomolecules. The biomolecules and cells/spores recovered from each collection device were assessed by cultivable and microscopic enumeration, and quantitative and species-specific PCR assays. rRNA gene-based quantitative PCR analysis showed that cotton swabs were superior to nylon-flocked swabs for sampling of small surface areas, and for larger surfaces, biological sampling kits significantly outperformed polyester wipes. Species-specific PCR revealed differential recovery of certain species dependent upon the sampling device employed. The results of this study empower current and future molecular-analysis-based microbial sampling and processing methodologies.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | SPACECRAFT ASSEMBLY FACILITY; BACILLUS-ANTHRACIS SPORES; NONPOROUS SURFACES; CLEAN ROOMS; SWAB PROTOCOL; MICROBIAL-CONTAMINATION; RECOVERY; DIVERSITY; BACTERIA; DNA; |
| Subjects: | 500 Science > 570 Life sciences |
| Divisions: | Biology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie > Lehrstuhl für Mikrobiologie (Archaeenzentrum) |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 22 Jun 2020 06:30 |
| Last Modified: | 22 Jun 2020 06:30 |
| URI: | https://pred.uni-regensburg.de/id/eprint/20892 |
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