Multiplex PCR for detection of plasmid-mediated quinolone resistance qnr genes in ESBL-producing enterobacterial isolates
Multiplex PCR for detection of plasmid-mediated quinolone resistance qnr genes in ESBL-producing enterobacterial isolates
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DOI：
10.1093/jac/dkm204
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发表时间：
2007-08-01
影响因子：
5.2
通讯作者：
Nordmann, Patrice
中科院分区：
文献类型：
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作者：
Cattoir, Vincent;Poirel, Laurent;Nordmann, Patrice
目的：目的：建立一种快速、可靠的单管PCR技术，同时检测质粒介导的喹诺酮类耐药基因qnrA、qnrB和qnrS。它们用于筛选2002年至2004年从科威特收集的64株产超广谱β-内酰胺酶（ESBL）的肠细菌分离株，因为ESBL基因通常与qnr基因相关。结果：在优化条件下，所有阳性对照（单独或混合使用）均证实了PCR引物的特异性。在64株分离株中，只有3株qnrrB样基因阳性（4.7%），而未检测到qnrA样基因和qnrS样基因。在阴沟肠杆菌K34（SHV-12+）分离株中检测到qnrB 2基因，而在大肠杆菌中检测到qnrB 1样基因（称为qnrB 7）和qnrB 6样基因（称为qnrB 8）。结论：本研究提供了一种快速筛选qnr阳性菌株的方法，可用于临床流行病学调查。在科威特分离株的研究中，确定了产ESBL肠杆菌科中Qnr决定簇的低流行率。
Objectives: To develop a rapid and reliable single-tube-based PCR technique for detecting simultaneously the plasmid-mediated quinolone resistance qnrA, qnrB and qnrS genes.Methods: After multiple alignments, primers were designed to detect known qnr variants (six for qnrA-, six for qnrB- and two for qnrS-like genes). They were used for screening a collection of 64 expanded-spectrum beta-lactamase (ESBL)-producing enterobacterial isolates from Kuwait, collected from 2002 to 2004, as ESBL genes have been often associated with qnr genes. Sequencing was performed to identify qnr and associated ESBL genes.Results: In optimized conditions, all positive controls (used separately or mixed) confirmed the specificity of the PCR primers. Out of 64 isolates, only 3 isolates were positive for a qnrrB-like gene (4.7%), whereas no qnrA-like and qnrS-like gene was detected. A qnrB2 gene was detected in an Enterobacter cloacae K34 (SHV-12+) isolate, whereas qnrB1-like (termed qnrB7) and qnrB6-like (termed qnrB8) genes were identified from E. cloacae K37 (SHV-12 +) and Citrobacter freundii K70 (VEB-1b +) isolates, respectively.Conclusions: We report here a fast and reliable technique for rapid screening of qnr-positive strains to be used for epidemiological surveys. A low prevalence of Qnr determinants among ESBL-producing Enterobacteriaceae was identified in the study with Kuwaiti isolates.