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首页> 外文期刊>Environmental Science & Technology >Development of Microbial Genome-Probing Microarrays Using Digital Multiple Displacement Amplification of Uncultivated Microbial Single Cells
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Development of Microbial Genome-Probing Microarrays Using Digital Multiple Displacement Amplification of Uncultivated Microbial Single Cells

机译：利用未经培养的微生物单细胞的数字多位移扩增技术开发微生物基因组探测芯片

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摘要

A crucial problem in the use of previously developed genome-probing microarrays (GPM) has been the inability to use uncultivated bacterial genomes to take advantage of the high sensitivity and specificity of GPM in microbial detection and monitoring. We show here a method, digital multiple displacement amplification (MDA), to amplify and analyze various genomes obtained from single uncultivated bacterial cells. We used 15 genomes from key microbes involved in dichloromethane (DCM)-dechlorinating enrichment as microarray probes to uncover the bacterial population dynamics of samples without PCR amplification. Genomic DNA amplified from single cells originating from uncultured bacteria with 80.3-99.4% similarity to 16S rRNA genes of cultivated bacteria. The digital MDA-GPM method successfully monitored the dynamics of DCM-dechlorinating communities from different phases of enrichment status. Without a priori knowledge of microbial diversity, the digital MDA-GPM method could be designed to monitor most microbial populations in a given environmental sample.
机译：使用先前开发的基因组探测微阵列（GPM）的一个关键问题是无法使用未经培养的细菌基因组来利用GPM在微生物检测和监测中的高灵敏度和特异性。我们在这里显示了一种方法，数字多位移扩增（MDA），以扩增和分析从单个未培养细菌细胞获得的各种基因组。我们使用了涉及二氯甲烷（DCM）脱氯富集的关键微生物的15个基因组作为微阵列探针来发现未经PCR扩增的样品的细菌种群动态。从源自未培养细菌的单细胞扩增基因组DNA，与培养细菌的16S rRNA基因有80.3-99.4％的相似性。数字MDA-GPM方法成功地监测了富集状态不同阶段的DCM-脱氯群落的动态。在没有先验微生物多样性知识的情况下，可以将数字MDA-GPM方法设计为监视给定环境样本中的大多数微生物种群。

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