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Metagenomics-Based Proficiency Test of Smoked Salmon Spiked with a Mock Community

机译:烟熏三文鱼的烟草术基水平试验飙升杂碎的群落

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摘要

An inter-laboratory proficiency test was organized to assess the ability of participants to perform shotgun metagenomic sequencing of cold smoked salmon, experimentally spiked with a mock community composed of six bacteria, one parasite, one yeast, one DNA, and two RNA viruses. Each participant applied its in-house wet-lab workflow(s) to obtain the metagenomic dataset(s), which were then collected and analyzed using MG-RAST. A total of 27 datasets were analyzed. Sample pre-processing, DNA extraction protocol, library preparation kit, and sequencing platform, influenced the abundance of specific microorganisms of the mock community. Our results highlight that despite differences in wet-lab protocols, the reads corresponding to the mock community members spiked in the cold smoked salmon, were both detected and quantified in terms of relative abundance, in the metagenomic datasets, proving the suitability of shotgun metagenomic sequencing as a genomic tool to detect microorganisms belonging to different domains in the same food matrix. The implementation of standardized wet-lab protocols would highly facilitate the comparability of shotgun metagenomic sequencing dataset across laboratories and sectors. Moreover, there is a need for clearly defining a sequencing reads threshold, to consider pathogens as detected or undetected in a food sample.
机译:组织了一个实验室熟练程度测试,以评估参与者对感冒熏鲑鱼进行霰弹枪梅毒测序的能力,通过由六个细菌,一种寄生虫,一种酵母,一种DNA和两个RNA病毒组成的模拟界进行了实验飙升。每个参与者将其内部湿式实验室工作流程应用于获得偏见数据集,然后使用Mg-Rast收集和分析。共分析了27个数据集。样品预处理,DNA提取方案,文库制备试剂盒和测序平台,影响了模拟群落的特异性微生物的丰富。我们的结果强调,尽管湿卫生实验室协议存在差异,但对与冷烟熏三文鱼中尖刺的模拟群落成员相对应的读数均以在肉桂元数据集中的相对丰度方面检测和量化,证明了霰弹枪偏心测序的适用性作为一种基因组工具,以检测属于同一食物基质中不同域的微生物。规范化的湿式实验室协议的实施将高度促进霰弹枪偏心序列测序数据集的可比性,跨实验室和部门。此外,需要清楚地定义测序读取阈值,以考虑如食物样本中检测到或未检测到的病原体。

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