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Evaluation of Procedures for the Collection, Processing, and Analysis of Biomolecules from Low-Biomass Surfaces

机译：评价从低生物量表面收集，加工和分析生物分子的程序

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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.
机译：为了通过基于分子分析的方法全面评估微生物的多样性和丰度，对样品收集，处理和分析的程序进行了深入评估。已知组成的模型微生物群落（MMC）（代表典型的低生物量表面样品）用于检查变量在采样矩阵，目标细胞密度/分子浓度和低温存储中对采样总体功效的影响。养生。在这项研究中使用的MMC包含11种与航天器或洁净室表面相关的细菌，古细菌和真菌谱系。将已知细胞密度的MMC沉积在不锈钢样板上，干燥后，使用各种采样设备回收细胞和生物分子。从每个收集设备中回收的生物分子和细胞/孢子通过可培养和微观计数以及定量和物种特异性PCR分析进行评估。基于rRNA基因的定量PCR分析表明，棉签在小表面积采样方面优于尼龙植签，对于较大的表面积，生物采样套件的性能明显优于聚酯抹布。物种特异性PCR显示，某些物种的差异回收取决于所采用的采样设备。这项研究的结果使当前和未来的基于分子分析的微生物采样和处理方法成为可能。

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