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在产酸克雷伯菌 M19 中,碳青霉烯类耐药基因 bla 由含有新型转座子 Tn6681 的可接合质粒传播。

The carbapenem resistance gene bla is disseminated by a conjugative plasmid containing the novel transposon Tn6681 in Acinetobacter johnsonii M19.

机构信息

Key Laboratory of Industrial Biotechnology of Ministry of Education and School of Biotechnology, Jiangnan University, Wuxi, 214122, China.

Department of Epidemiology, The First Affiliated Hospital of Shandong First Medical University, Jinan, 250062, China.

出版信息

Antimicrob Resist Infect Control. 2020 Nov 9;9(1):182. doi: 10.1186/s13756-020-00832-4.

Abstract

BACKGROUND

Carbapenem resistant Acinetobacter species have caused great difficulties in clinical therapy in the worldwide. Here we describe an Acinetobacter johnsonii M19 with a novel bla containing transposon Tn6681 on the conjugative plasmid pFM-M19 and the ability to transferand carbapenem resistance.

METHODS

A. johnsonii M19 was isolated under selection with 8 mg/L meropenem from hospital sewage, and the minimum inhibitory concentrations (MICs) for the representative carbapenems imipenem, meropenem and ertapenem were determined. The genome of A. johnsonii M19 was sequenced by PacBio RS II and Illumina HiSeq 4000 platforms. A homologous model of OXA-23 was generated, and molecular docking models with imipenem, meropenem and ertapenem were constructed by Discovery Studio 2.0. Type IV secretion system and conjugation elements were identified by the Pathosystems Resource Integration Center (PATRIC) server and the oriTfinder. Mating experiments were performed to evaluate transfer of OXA-23 to Escherichia coli 25DN.

RESULTS

MICs of A. johnsonii M19 for imipenem, meropenem and ertapenem were 128 mg/L, 48 mg/L and 24 mg/L, respectively. Genome sequencing identified plasmid pFM-M19, which harbours the carbapenem resistance gene bla within the novel transposon Tn6681. Molecular docking analysis indicated that the elongated hydrophobic tunnel of OXA-23 provides a hydrophobic environment and that Lys-216, Thr-217, Met-221 and Arg-259 were the conserved amino acids bound to imipenem, meropenem and ertapenem. Furthermore, pFM-M19 could transfer bla to E. coli 25DN by conjugation, resulting in carbapenem-resistant transconjugants.

CONCLUSIONS

Our investigation showed that A. johnsonii M19 is a source and disseminator of bla and carbapenem resistance. The ability to transfer bla to other species by the conjugative plasmid pFM-M19 raises the risk of spread of carbapenem resistance. The carbapenem resistance gene bla is disseminated by a conjugative plasmid containing the novel transposon Tn6681 in Acinetobacter johnsonii M19.

摘要

背景

耐碳青霉烯不动杆菌属在全球范围内的临床治疗中造成了极大的困难。在这里,我们描述了一株携带新型 bla 的约翰逊不动杆菌 M19,该 bla 位于可转移的质粒 pFM-M19 上,并具有携带和碳青霉烯类药物耐药性的能力。

方法

从医院污水中选择 8mg/L 美罗培南,分离出约翰逊不动杆菌 M19,测定代表碳青霉烯类药物亚胺培南、美罗培南和厄他培南的最小抑菌浓度(MIC)。使用 PacBio RS II 和 Illumina HiSeq 4000 平台对约翰逊不动杆菌 M19 的基因组进行测序。生成 OXA-23 的同源模型,并通过 Discovery Studio 2.0 构建与亚胺培南、美罗培南和厄他培南的分子对接模型。通过 Pathosystems Resource Integration Center(PATRIC)服务器和 oriTfinder 鉴定了 IV 型分泌系统和接合元件。进行交配实验以评估 OXA-23 向大肠杆菌 25DN 的转移。

结果

约翰逊不动杆菌 M19 对亚胺培南、美罗培南和厄他培南的 MIC 分别为 128mg/L、48mg/L 和 24mg/L。基因组测序鉴定出质粒 pFM-M19,其携带 bla 基因位于新型转座子 Tn6681 内。分子对接分析表明,OXA-23 的长疏水性隧道提供了一个疏水性环境,Lys-216、Thr-217、Met-221 和 Arg-259 是与亚胺培南、美罗培南和厄他培南结合的保守氨基酸。此外,pFM-M19 可以通过接合将 bla 转移到大肠杆菌 25DN,导致碳青霉烯类耐药的转导子。

结论

我们的研究表明,约翰逊不动杆菌 M19 是 bla 和碳青霉烯类药物耐药性的来源和传播者。可转移质粒 pFM-M19 将 bla 转移到其他物种的能力增加了碳青霉烯类耐药性传播的风险。碳青霉烯类耐药基因 bla 由约翰逊不动杆菌 M19 中的新型转座子 Tn6681 携带的可接合质粒传播。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d75d/7653874/4da32e0fa5db/13756_2020_832_Fig1_HTML.jpg

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