Wang Minghua, Guo Qinglan, Xu Xiaogang, Wang Xiaoying, Ye Xinyu, Wu Shi, Hooper David C, Wang Minggui
Institute of Antibiotics, Huashan Hospital, Fudan University, 12 M. Wulumuqi Rd., Shanghai 200040, People's Republic of China.
Antimicrob Agents Chemother. 2009 May;53(5):1892-7. doi: 10.1128/AAC.01400-08. Epub 2009 Mar 2.
Since the discovery of qnrA in 1998, two additional qnr genes, qnrB and qnrS, have been described. These three plasmid-mediated genes contribute to quinolone resistance in gram-negative pathogens worldwide. A clinical strain of Proteus mirabilis was isolated from an outpatient with a urinary tract infection and was susceptible to most antimicrobials but resistant to ampicillin, sulfamethoxazole, and trimethoprim. Plasmid pHS10, harbored by this strain, was transferred to azide-resistant Escherichia coli J53 by conjugation. A transconjugant with pHS10 had low-level quinolone resistance but was negative by PCR for the known qnr genes, aac(6')-Ib-cr and qepA. The ciprofloxacin MIC for the clinical strain and a J53/pHS10 transconjugant was 0.25 microg/ml, representing an increase of 32-fold relative to that for the recipient, J53. The plasmid was digested with HindIII, and a 4.4-kb DNA fragment containing the new gene was cloned into pUC18 and transformed into E. coli TOP10. Sequencing showed that the responsible 666-bp gene, designated qnrC, encoded a 221-amino-acid protein, QnrC, which shared 64%, 42%, 59%, and 43% amino acid identity with QnrA1, QnrB1, QnrS1, and QnrD, respectively. Upstream of qnrC there existed a new IS3 family insertion sequence, ISPmi1, which encoded a frameshifted transposase. qnrC could not be detected by PCR, however, in 2,020 strains of Enterobacteriaceae. A new quinolone resistance gene, qnrC, was thus characterized from plasmid pHS10 carried by a clinical isolate of P. mirabilis.
自1998年发现qnrA以来,又描述了另外两个qnr基因,即qnrB和qnrS。这三个质粒介导的基因致使全球范围内革兰氏阴性病原体对喹诺酮产生耐药性。从一名患有尿路感染的门诊患者中分离出一株奇异变形杆菌临床菌株,该菌株对大多数抗菌药物敏感,但对氨苄西林、磺胺甲恶唑和甲氧苄啶耐药。该菌株携带的质粒pHS10通过接合转移至叠氮化物抗性大肠杆菌J53。携带pHS10的接合子具有低水平喹诺酮耐药性,但针对已知的qnr基因、aac(6')-Ib-cr和qepA的PCR检测呈阴性。临床菌株和J53/pHS10接合子的环丙沙星MIC为0.25μg/ml,相对于受体J53增加了32倍。用HindIII消化该质粒,将包含新基因的4.4kb DNA片段克隆到pUC18中并转化到大肠杆菌TOP10中。测序表明,负责的666bp基因(命名为qnrC)编码一种221个氨基酸的蛋白质QnrC,其与QnrA1、QnrB1、QnrS1和QnrD的氨基酸同一性分别为64%、42%、59%和43%。在qnrC上游存在一个新的IS3家族插入序列ISPmi1,其编码一个移码转座酶。然而,在2020株肠杆菌科菌株中通过PCR未检测到qnrC。因此,从奇异变形杆菌临床分离株携带的质粒pHS10中鉴定出一个新的喹诺酮耐药基因qnrC。