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产超广谱β-内酰胺酶大肠埃希菌对上皮细胞的黏附与侵袭及运动性显示ST131优势:肠外致病性大肠埃希菌谱系的体外比较研究

Adhesion and invasion to epithelial cells and motility of extended-spectrum β-lactamase-producing Escherichia coli reveal ST131 superiority: a comparative in vitro study of extraintestinal pathogenic E. coli lineages.

作者信息

Kondratyeva Kira, Wollman Ayala, Gerlitz Gabi, Navon-Venezia Shiri

机构信息

Department of Molecular Biology, Faculty of Life Science, Ariel University, Ariel, Israel.

出版信息

J Med Microbiol. 2017 Sep;66(9):1350-1357. doi: 10.1099/jmm.0.000549. Epub 2017 Aug 22.

Abstract

PURPOSE

Extended-spectrum β-lactamase (ESBL)-producing extraintestinal pathogenic Escherichia coli (ExPEC) sequence type ST131 is pandemic, and it is the major contributor to antibiotic resistance in E. coli. Despite its epidemiological superiority, the physiological reasons that decipher its success remain elusive. We aimed to compare the adhesion, invasion and motility potential of ST131 versus other E. coli lineages.

METHODOLOGY

In this in vitro comparative study, 14 ESBL-producing ExPEC community-onset bacteremia isolates were chosen from a reported clinical collection (Karfunkel D, Carmeli Y, Chmelnitsky I, Kotlovsky T, Navon-Venezia S. Eur J Clin Microbiol Infect Dis 2013;32:513-521). Isolates were divided into two groups, ST131 (n=7) and 'non-ST131', sporadic sequence types (STs) (n=7). Virulence and adhesion genes were screened by PCR in all isolates. Virotyping and serotyping were performed for ST131 isolates. Adhesion and invasion to Caco-2 epithelial cells, and motility on semi-solid agar were quantified and compared between the two groups. Fluorescence microscopy using anti-LPS E. coli antibodies was used for visualization and confirmation of adhesion and invasion.

RESULTS

ST131 isolates belonged to the O25b:H4-B2 subclone. Two ST131 virotypes were found, A (two blaCTX-M-15 H30-Rx) and C (two blaCTX-M-15 H30-Rx and three blaCTX-M-14 H30 isolates). The average number of adhesion and virulence genes carried by ExPEC ST131 isolates and non-ST131 isolates was 5.3 and 3.7, respectively (P<0.05). Group analysis showed that ST131 surpassed non-ST131 lineages in all three physiological properties: adherence (17.1 vs 13.1 %, P<0.001), invasion (0.4 vs 0.17 %, P<0.01), and swarming motility on all media tested (P<0.05).

CONCLUSION

This study demonstrates ST131 superiority that may explain its improved gut-colonization and dissemination capabilities within the host. These insights are an important step in our understanding of ST131 epidemiological success.

摘要

目的

产超广谱β-内酰胺酶(ESBL)的肠外致病性大肠杆菌(ExPEC)序列类型ST131呈全球流行,是大肠杆菌抗生素耐药性的主要成因。尽管其在流行病学上具有优势,但其成功的生理原因仍不清楚。我们旨在比较ST131与其他大肠杆菌谱系的黏附、侵袭和运动能力。

方法

在这项体外比较研究中,从已报道的临床样本中选取了14株产ESBL的ExPEC社区获得性菌血症分离株(Karfunkel D, Carmeli Y, Chmelnitsky I, Kotlovsky T, Navon-Venezia S. Eur J Clin Microbiol Infect Dis 2013;32:513 - 521)。分离株分为两组,ST131组(n = 7)和“非ST131”散发性序列类型(STs)组(n = 7)。通过PCR对所有分离株的毒力和黏附基因进行筛查。对ST131分离株进行病毒分型和血清分型。对两组之间黏附并侵袭Caco - 2上皮细胞的情况以及在半固体琼脂上的运动能力进行定量和比较。使用抗LPS大肠杆菌抗体的荧光显微镜用于观察和确认黏附与侵袭情况。

结果

ST131分离株属于O25b:H4 - B2亚克隆。发现了两种ST131病毒分型,A(两个blaCTX - M - 15 H30 - Rx)和C(两个blaCTX - M - 15 H30 - Rx以及三个blaCTX - M - 14 H30分离株)。ExPEC ST131分离株和非ST131分离株携带的黏附基因和毒力基因的平均数分别为5.3和3.7(P < 0.05)。分组分析表明,ST131在所有三种生理特性上均超过非ST131谱系:黏附(17.1%对13.1%,P < 0.001)、侵袭(0.4%对0.17%,P < 0.01)以及在所有测试培养基上的群集运动能力(P < 0.05)。

结论

本研究证明了ST131的优势,这可能解释了其在宿主体内改善的肠道定植和传播能力。这些见解是我们理解ST131流行病学成功的重要一步。

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