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[大肠杆菌菌株中质粒介导的喹诺酮耐药性研究]

[Investigation of plasmid-mediated quinolone resistance in Escherichia coli strains].

作者信息

Aktepe Orhan Cem, Aşık Gülşah, Cetinkol Yeliz, Biçmen Meral, Gülay Zeynep

机构信息

Afyon Kocatepe University Faculty of Medicine, Department of Medical Microbiology, Afyonkarahisar, Turkey.

出版信息

Mikrobiyol Bul. 2012 Jan;46(1):9-16.

Abstract

Quinolones are widely used antimicrobial agents, particularly for the treatment of infections caused by gram-negative bacilli such as E.coli. As a consequence, quinolone resistance has been increasing among this species in recent years. Bacterial resistance to quinolones usually results from mutations in the chromosomal genes which encode topoisomerases and also the expression of efflux pumps and loss of porines contributed to development of quinolone resistance. However, recent studies have shown that the spread and increase of quinolone resistance may be due to the transfer of plasmid-mediated genes. To date, three groups of plasmid-mediated quinolone resistance genes, namely qnr, aac(6')-Ib-cr, and qepA, have been described. The aim of this study was to investigate the presence of plasmid-mediated quinolone resistance genes in E.coli clinical isolates. A total of 112 quinolone-resistant E.coli strains isolated from different clinical specimens (84 urine, 16 blood, 10 wound, 2 bronchoalveolar lavage) of which 78 (69.6%) were extended-spectrum beta-lactamase (ESBL) positive, in Afyon Kocatepe University Hospital, Microbiology Laboratory were included in the study. In the isolates, qnrA, qnrB, qnrS, qnrC, qepA, and aac(6')-1b-cr plasmid genes were analysed by polymerase chain reaction (PCR). After aac(6')- 1b determinant was amplified by PCR, all aac(6')-1b positive amplicons were analyzed by digestion with BseGI restriction enzyme to identify aac(6')-1b-cr variant. It was found that, none of the strains horboured qnrA, qnrB, qnrS, qnrC and qepA genes, however, plasmid-mediated quinolone resistance gene aac(6')-1b-cr was found positive in 59.8% (67/112) of the strains. It was notable that 86.6% (58/67) of those isolates were ESBL producers. The rates of quinolone resistance among E.coli isolates infections were high in our region and an increasing trend has been observed in recent years. Our data indicated that the presence of plasmid- mediated resistance genes such as aac(6')-1b-cr, might have contributed to the high quinolone resistance rates. In conclusion, not only qnr genes but all other plasmid-mediated quinolone resistance genes, should be tested for the detection of plasmid-mediated quinolone resistance and this fact should be taken into consideration when the reservoirs are being searched for.

摘要

喹诺酮类是广泛使用的抗菌药物,尤其用于治疗由革兰氏阴性杆菌如大肠杆菌引起的感染。因此,近年来该菌属中喹诺酮耐药性一直在增加。细菌对喹诺酮类的耐药性通常源于编码拓扑异构酶的染色体基因突变,以及外排泵的表达和孔蛋白的缺失,这些都导致了喹诺酮耐药性的产生。然而,最近的研究表明,喹诺酮耐药性的传播和增加可能是由于质粒介导基因的转移。迄今为止,已描述了三组质粒介导的喹诺酮耐药基因,即qnr、aac(6')-Ib-cr和qepA。本研究的目的是调查大肠杆菌临床分离株中质粒介导的喹诺酮耐药基因的存在情况。在阿菲永科贾泰佩大学医院微生物实验室,共纳入了112株从不同临床标本(84份尿液、16份血液、10份伤口标本、2份支气管肺泡灌洗标本)分离出的喹诺酮耐药大肠杆菌菌株,其中78株(69.6%)超广谱β-内酰胺酶(ESBL)阳性。对分离株中的qnrA、qnrB、qnrS、qnrC、qepA和aac(6')-1b-cr质粒基因进行聚合酶链反应(PCR)分析。通过PCR扩增aac(6')- 1b决定簇后,所有aac(6')-1b阳性扩增子用BseGI限制酶消化进行分析,以鉴定aac(6')-1b-cr变体。结果发现,所有菌株均未携带qnrA、qnrB、qnrS、qnrC和qepA基因,然而,质粒介导的喹诺酮耐药基因aac(6')-1b-cr在59.8%(67/112)的菌株中呈阳性。值得注意的是,这些分离株中有86.6%(58/67)是ESBL产生菌。在我们地区,大肠杆菌分离株感染中的喹诺酮耐药率很高,且近年来呈上升趋势。我们的数据表明,aac(6')-1b-cr等质粒介导的耐药基因的存在可能导致了高喹诺酮耐药率。总之,为了检测质粒介导的喹诺酮耐药性,不仅应检测qnr基因,还应检测所有其他质粒介导的喹诺酮耐药基因,并且在寻找储存库时应考虑到这一事实。

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