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首页> 外文期刊>Journal of Clinical Microbiology >Direct 16S rRNA Gene Sequencing from Clinical Specimens, with Special Focus on Polybacterial Samples and Interpretation of Mixed DNA Chromatograms
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Direct 16S rRNA Gene Sequencing from Clinical Specimens, with Special Focus on Polybacterial Samples and Interpretation of Mixed DNA Chromatograms

机译：从临床标本中直接进行16S rRNA基因测序，重点关注多细菌样品和混合DNA色谱图的解释

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摘要

RipSeq (iSentio, Bergen, Norway) is a web-based application for the analysis of mixed DNA chromatograms. It opens the possibility to analyze chromatograms obtained by direct 16S rRNA gene sequencing from polybacterial human clinical samples. In this study, we used direct 16S rRNA gene sequencing to investigate 264 samples from a wide range of suspected human bacterial infections. The sequence-based identification was compared with the results from routine culture-based identification. A total of 151 samples were positive by the first PCR, producing 85 pure and 66 mixed DNA chromatograms. All mixed chromatograms were analyzed by RipSeq, although seven were so complex that only the dominant bacterial sequences could be identified. In general, sequence-based identification detected a larger number of species than did culture for samples from patients who had received antibiotics prior to sample collection and for samples containing anaerobic bacteria. RipSeq made it possible to apply this supplementary diagnostic tool to typical polybacterial specimens, such as internal abscesses, pleural fluids, and bile.
机译：RipSeq（iSentio，卑尔根，挪威）是一种基于Web的应用程序，用于分析混合DNA色谱图。这为分析通过16S rRNA基因直接测序从多细菌人类临床样品中获得的色谱图提供了可能性。在这项研究中，我们使用直接16S rRNA基因测序来调查来自广泛的人类细菌感染的264个样本。将基于序列的鉴定与基于常规培养的鉴定的结果进行了比较。第一次PCR共有151个样品呈阳性，产生了85个纯净色谱图和66个混合DNA色谱图。所有混合色谱图都通过RipSeq进行了分析，尽管其中有7个非常复杂，以至于只能识别出主要的细菌序列。一般而言，基于序列的鉴定所检测到的物种数量要比培养之前为收集样本之前已接受抗生素的患者的样本以及含有厌氧细菌的样本所检测的物种数量多。 RipSeq使得可以将这种辅助诊断工具应用于典型的多细菌标本，例如内部脓肿，胸腔积液和胆汁。

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