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Evaluating the mobility potential of antibiotic resistance genes in environmental resistomes without metagenomics

机译:在没有宏基因组学的情况下评估抗生素抗性基因在环境抗药性中的迁移潜力

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摘要

Antibiotic resistance genes are ubiquitous in the environment. However, only a fraction of them are mobile and able to spread to pathogenic bacteria. Until now, studying the mobility of antibiotic resistance genes in environmental resistomes has been challenging due to inadequate sensitivity and difficulties in contig assembly of metagenome based methods. We developed a new cost and labor efficient method based on Inverse PCR and long read sequencing for studying mobility potential of environmental resistance genes. We applied Inverse PCR on sediment samples and identified 79 different MGE clusters associated with the studied resistance genes, including novel mobile genetic elements, co-selected resistance genes and a new putative antibiotic resistance gene. The results show that the method can be used in antibiotic resistance early warning systems. In comparison to metagenomics, Inverse PCR was markedly more sensitive and provided more data on resistance gene mobility and co-selected resistances.
机译:抗生素抗性基因在环境中无处不在。但是,它们中只有一小部分是可移动的并且能够传播到病原细菌。迄今为止,由于敏感性不足和基于元基因组的重叠群组装中的困难,研究环境抵抗组中抗生素抗性基因的迁移一直具有挑战性。我们开发了一种基于反向PCR和长读测序的新方法,具有成本效益和劳动效率,用于研究环境抗性基因的迁移潜力。我们对沉积物样品进行了反向PCR,发现了与研究的抗性基因相关的79个不同的MGE簇,包括新型的流动遗传元件,共同选择的抗性基因和新的推定的抗生素抗性基因。结果表明,该方法可用于抗生素耐药性预警系统。与宏基因组学相比,反向PCR的敏感性更高,并提供了有关抗性基因迁移性和共选抗性的更多数据。

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