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检测动物源大肠杆菌分离株中的 qnr 基因和携带 qnrB19 的可接合质粒 pQNR2078 的完整序列。

Detection of qnr genes among Escherichia coli isolates of animal origin and complete sequence of the conjugative qnrB19-carrying plasmid pQNR2078.

机构信息

Institute of Farm Animal Genetics, Friedrich-Loeffler-Institut, Neustadt-Mariensee, Germany.

出版信息

J Antimicrob Chemother. 2012 May;67(5):1099-102. doi: 10.1093/jac/dks024. Epub 2012 Feb 14.

Abstract

OBJECTIVES

The aims of this study were to identify qnr genes among quinolone-resistant Escherichia coli isolates from defined disease conditions of companion and farm animals obtained in the BfT-GermVet study, and to gain insight into their localization and the organization of the qnr gene regions.

METHODS

The qnr genes were detected by PCR and confirmed by sequencing. qnr-positive isolates were checked for mutations in DNA gyrase and topoisomerase IV genes by PCR and sequencing of the quinolone resistance-determining regions. Multilocus sequence typing (MLST) was performed for the qnr-positive E. coli isolates. Plasmids harbouring qnr genes were transferred by conjugation into E. coli recipients, subjected to PCR-based replicon typing and plasmid-MLST, and one qnrB19-carrying plasmid was sequenced completely.

RESULTS

Only 2 of 417 E. coli isolates investigated carried qnr genes. Both isolates originated from horses and showed MLST type ST86. They harboured conjugative qnrB19-carrying plasmids, which proved to be indistinguishable by restriction analysis, belonged to incompatibility group IncN, showed plasmid-MLST type ST8 and did not carry other resistance genes. The qnrB19 gene was flanked by copies of the insertion sequence IS26. One of these plasmids, pQNR2078, was sequenced completely and had a size of 42 379 bp. Except for the resistance gene region, plasmid pQNR2078 closely resembled the bla(CTX-M-65)-carrying plasmid pKC396 from E. coli.

CONCLUSIONS

qnr genes were rarely detected among E. coli from animals in the BfT-GermVet study. The qnrB19 gene was detected on conjugative plasmids, with IS26 being likely involved in the mobility of qnrB19.

摘要

目的

本研究旨在鉴定 BfT-GermVet 研究中从伴侣动物和农场动物特定疾病分离的耐喹诺酮大肠埃希菌中 qnr 基因,并深入了解其定位和 qnr 基因区域的组织。

方法

通过 PCR 检测 qnr 基因,并通过测序进行确认。qnr 阳性分离株通过 PCR 和喹诺酮耐药决定区的测序检查 DNA 拓扑异构酶和拓扑异构酶 IV 基因突变。对 qnr 阳性大肠埃希菌分离株进行多位点序列分型(MLST)。通过接合将携带 qnr 基因的质粒转移到大肠埃希菌受体中,进行基于 PCR 的复制子分型和质粒-MLST,并对一个携带 qnrB19 的质粒进行完全测序。

结果

在所研究的 417 株大肠埃希菌分离株中,只有 2 株携带 qnr 基因。这两个分离株均来自马,其 MLST 型为 ST86。它们携带可接合的 qnrB19 携带质粒,通过限制分析证明无法区分,属于不相容群 IncN,质粒-MLST 型为 ST8,不携带其他耐药基因。qnrB19 基因两侧是插入序列 IS26 的拷贝。其中一个质粒,pQNR2078,被完全测序,大小为 42379bp。除了耐药基因区域外,质粒 pQNR2078 与大肠埃希菌中携带 bla(CTX-M-65)的质粒 pKC396 非常相似。

结论

BfT-GermVet 研究中从动物分离的大肠埃希菌中很少检测到 qnr 基因。qnrB19 基因位于可接合的质粒上,IS26 可能参与了 qnrB19 的移动。

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