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从人类粪便中分离并从临床细菌分离物中诱导的噬菌体颗粒中的抗生素耐药基因。

Antibiotic resistance genes in phage particles isolated from human faeces and induced from clinical bacterial isolates.

机构信息

Department of Genetics, Microbiology and Statistics, University of Barcelona, Diagonal 643, Annex, Floor 0, E-08028 Barcelona, Spain.

Servei de Microbiologia, Hospital de la Santa Creu i Sant Pau, Institut d'Investigació Biomèdica Sant Pau, Sant Quintí 89, 08041 Barcelona, Spain.

出版信息

Int J Antimicrob Agents. 2018 Mar;51(3):434-442. doi: 10.1016/j.ijantimicag.2017.11.014. Epub 2017 Nov 24.

Abstract

Phage particles have emerged as elements with the potential to mobilise antibiotic resistance genes (ARGs) in different environments, including the intestinal habitat. This study aimed to determine the occurrence of ARGs in phage particles present in faecal matter and induced from strains isolated from faeces. Nine ARGs (bla, bla, bla, bla, qnrA, qnrS, mecA, sul1 and armA) were quantified by qPCR in the phage DNA fractions of 150 faecal samples obtained from healthy individuals who had not received antibiotic treatment or travelled abroad in the 3 months prior to sample collection. On the suspicion that the detected particles originated from bacterial flora, 82 Escherichia coli and Klebsiella pneumoniae isolates possessing at least one identified ARG (bla, bla, bla, armA, qnrA, qnrS and sul1) were isolated and their capacity to produce phage particles carrying these ARGs following induction was evaluated. Of 150 samples, 72.7% were positive for at least one ARG, with bla and bla being the most prevalent and abundant. Of the 82 isolates, 51 (62%) showed an increase in the number of copies of the respective ARG in the phage fraction following induction, with bla, bla, bla and sul1 being the most abundant. Phages induced from the isolates were further purified and visualised using microscopy and their DNA showed ARG levels of up to 10 gene copies/mL. This study highlights the abundance of phage particles harbouring ARGs and indicates that bacterial strains in the intestinal habitat could be source of these particles.

摘要

噬菌体颗粒已成为具有在不同环境中(包括肠道生境)移动抗生素耐药基因(ARGs)潜力的元素。本研究旨在确定粪便中存在的噬菌体颗粒中 ARGs 的发生情况,并从粪便中分离的菌株中诱导噬菌体颗粒。在未接受抗生素治疗或在采样前 3 个月内未出国旅行的 150 名健康个体的粪便样本中,通过 qPCR 定量了噬菌体 DNA 部分中的 9 个 ARGs(bla、bla、bla、bla、qnrA、qnrS、mecA、sul1 和 armA)。由于怀疑检测到的颗粒源自细菌菌群,因此分离了至少携带一种鉴定出的 ARG(bla、bla、bla、armA、qnrA、qnrS 和 sul1)的 82 株大肠杆菌和肺炎克雷伯菌,并评估了它们在诱导后产生携带这些 ARGs 的噬菌体颗粒的能力。在 150 个样本中,有 72.7%至少对一种 ARG 呈阳性,其中 bla 和 bla 最为普遍和丰富。在 82 株分离株中,有 51 株(62%)在诱导后噬菌体部分的相应 ARG 拷贝数增加,其中 bla、bla、bla 和 sul1 最为丰富。从分离株诱导的噬菌体进一步纯化并通过显微镜观察其 DNA,其 ARG 水平高达 10 个基因拷贝/ml。本研究强调了携带 ARGs 的噬菌体颗粒的丰富度,并表明肠道生境中的细菌菌株可能是这些颗粒的来源。

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