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美国肺炎克雷伯菌临床分离株中与qnr基因相关的新出现的质粒介导喹诺酮耐药性。

Emerging plasmid-mediated quinolone resistance associated with the qnr gene in Klebsiella pneumoniae clinical isolates in the United States.

作者信息

Wang Minggui, Sahm Daniel F, Jacoby George A, Hooper David C

机构信息

Division of Infectious Diseases, Massachusetts General Hospital, Boston, Massachusetts 02114, USA.

出版信息

Antimicrob Agents Chemother. 2004 Apr;48(4):1295-9. doi: 10.1128/AAC.48.4.1295-1299.2004.

Abstract

Although quinolone resistance commonly results from chromosomal mutation, recent studies indicate that such resistance can also be transferred on plasmids carrying the gene responsible, qnr. One hundred ten ciprofloxacin-resistant clinical isolates of Klebsiella pneumoniae and Escherichia coli from the United States were screened for the qnr gene by PCR and Southern hybridization of plasmid DNA. Conjugation experiments were done with azide-resistant E. coli J53 as the recipient and selection with azide and sulfonamide, a resistance frequently linked to qnr. EcoRI and BamHI digests of qnr-hybridizing plasmids were subjected to electrophoresis on agarose gels and probed with qnr by Southern hybridization. qnr was detected in 8 (11.1%) of 72 K. pneumoniae strains. These eight positive strains were from six states in the United States. qnr was not found in any of the 38 E. coli strains tested. Quinolone resistance was transferred from seven of the eight probe-positive strains. Transconjugants with qnr-hybridizing plasmids had 32-fold increases in ciprofloxacin MICs relative to E. coli J53. For all eight strains, the sequence of qnr was identical to that originally reported. By size and restriction digests, four plasmids were related to the first-reported plasmid, pMG252, and three were different. Five new qnr plasmids encoded FOX-5 beta-lactamase, as did pMG252, but two others produced SHV-7 extended-spectrum beta-lactamase. Transferable plasmid-mediated quinolone resistance associated with qnr is now widely distributed in quinolone-resistant clinical strains of K. pneumoniae in the United States. Plasmid-determined quinolone resistance contributes to the increasing quinolone resistance of K. pneumoniae isolates and to the linkage previously observed between resistance to quinolones and the latest beta-lactam antibiotics.

摘要

尽管喹诺酮耐药性通常源于染色体突变,但最近的研究表明,这种耐药性也可通过携带耐药基因qnr的质粒进行传递。通过PCR以及质粒DNA的Southern杂交,对来自美国的110株耐环丙沙星的肺炎克雷伯菌和大肠杆菌临床分离株进行了qnr基因筛查。以耐叠氮化物的大肠杆菌J53作为受体进行接合实验,并使用叠氮化物和磺胺进行筛选,磺胺耐药性常与qnr相关。对与qnr杂交的质粒进行EcoRI和BamHI酶切,然后在琼脂糖凝胶上进行电泳,并通过Southern杂交用qnr进行探针检测。在72株肺炎克雷伯菌菌株中有8株(11.1%)检测到qnr。这8株阳性菌株来自美国的6个州。在所检测的38株大肠杆菌菌株中均未发现qnr。喹诺酮耐药性从8株探针阳性菌株中的7株进行了转移。携带与qnr杂交质粒的接合子相对于大肠杆菌J53,其环丙沙星MIC增加了32倍。对于所有8株菌株,qnr的序列与最初报道的相同。根据大小和酶切分析,4种质粒与首次报道的质粒pMG252相关,另外3种则不同。5种新的qnr质粒编码FOX-5β-内酰胺酶,pMG252也是如此,但另外两种产生SHV-7超广谱β-内酰胺酶。与qnr相关的可转移质粒介导的喹诺酮耐药性目前在美国耐喹诺酮的肺炎克雷伯菌临床菌株中广泛分布。质粒介导的喹诺酮耐药性导致肺炎克雷伯菌分离株对喹诺酮的耐药性增加,并导致先前观察到的对喹诺酮类药物和最新β-内酰胺类抗生素耐药性之间的关联。

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