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从血流感染中分离出的产超广谱β-内酰胺酶大肠杆菌——抗菌药敏性、耐药基因的接合转移及系统发育起源

ESBL-producing Escherichia coli isolated from bloodstream infections--antimicrobial susceptibility, conjugative transfer of resistance genes and phylogenetic origin.

作者信息

Franiczek Roman, Krzyżanowska Barbara

机构信息

Department of Microbiology, Wroclaw Medical University, Poland.

出版信息

Adv Clin Exp Med. 2014 Nov-Dec;23(6):865-70. doi: 10.17219/acem/37328.

Abstract

BACKGROUND

The prevalence of bloodstream infections (BSIs) due to ESBL-producing Escherichia coli (ESBL-EC) strains has increased dramatically over the past years.

OBJECTIVES

Characterization of ESBL-EC isolates collected from BSIs with regard to their antimicrobial susceptibility and phylogenetic background. The conjugative transfer of resistance determinants to the E. coli reference strain K12 C600 was also investigated.

MATERIAL AND METHODS

A collection of forty-eight ESBL-EC strains recovered from BSIs was subjected to the study. These strains were obtained from the ICU (intensive care unit) of the Medical University Hospital, Wrocław, Poland, during a four-year period (2009-2012). All the isolates were screened for ESBL production by the double disk synergy test (DDST). Transferability of plasmid-mediated resistance genes was performed by the conjugational broth method. Susceptibility to antibiotics and chemotherapeutics of clinical isolates and transconjugants was determined by the agar dilution method. PCR assay was used to detect the blaCTX-M gene in ESBL-EC tested and transconjugants. Affiliation to phylogenetic groups was done by the triplex PCR method.

RESULTS

Conjugational transfer of plasmids responsible for ESBL to a recipient strain was successful for all the ESBL-EC analyzed (donors). The conjugation frequencies ranging from 2.3×10(-7) to 5.2×10(-1) per donor. In vitro susceptibility testing revealed that all the ESBL-EC isolates and their transconjugants were resistant to most of the antimicrobial agents tested with the exception of carbapenems, tigecycline, and β-lactam-clavulanate combinations. Moreover, all the donor strains and their transconjugants were found to contain the blaCTX-M gene. The majority of the isolates analyzed belonged to phylogroups B2 (62.5%) and D (25%), whereas groups B1 and A were less frequently represented (8.3% and 4.2%, respectively).

CONCLUSIONS

The results of the study confirm the need of antibiotic policies and effective infection control measures in hospital settings to minimize BSIs caused by multi-resistant ESBL-producing pathogens.

摘要

背景

在过去几年中,产超广谱β-内酰胺酶(ESBL)的大肠埃希菌(ESBL-EC)引起的血流感染(BSIs)患病率急剧上升。

目的

对从BSIs中分离出的ESBL-EC菌株进行抗菌药敏性和系统发育背景的特征分析。同时还研究了耐药决定因子向大肠埃希菌参考菌株K12 C600的接合转移情况。

材料与方法

对从BSIs中分离出的48株ESBL-EC菌株进行研究。这些菌株是在四年期间(2009 - 2012年)从波兰弗罗茨瓦夫医科大学医院的重症监护病房(ICU)获得的。通过双纸片协同试验(DDST)对所有分离株进行ESBL产生的筛选。采用接合肉汤法检测质粒介导的耐药基因的可转移性。通过琼脂稀释法测定临床分离株和接合子对抗生素及化疗药物的敏感性。采用PCR检测法检测受试ESBL-EC菌株和接合子中的blaCTX-M基因。通过三重PCR法确定其所属的系统发育群。

结果

对所有分析的ESBL-EC(供体)菌株而言,负责ESBL的质粒向受体菌株的接合转移均成功。每个供体的接合频率范围为2.3×10(-7)至5.2×10(-1)。体外药敏试验表明,除碳青霉烯类、替加环素和β-内酰胺 - 克拉维酸联合制剂外,所有ESBL-EC分离株及其接合子对大多数受试抗菌药物耐药。此外,所有供体菌株及其接合子均被发现含有blaCTX-M基因。分析的大多数分离株属于系统发育群B2(62.5%)和D(25%),而B1群和A群的代表较少(分别为8.3%和4.2%)。

结论

研究结果证实,医院环境中需要制定抗生素政策和有效的感染控制措施,以尽量减少由多重耐药的产ESBL病原体引起的BSIs。

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