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首页> 外文期刊>The Journal of Antimicrobial Chemotherapy >Multiplex PCR for detection of plasmid-mediated quinolone resistance qnr genes in ESBL-producing enterobacterial isolates.
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Multiplex PCR for detection of plasmid-mediated quinolone resistance qnr genes in ESBL-producing enterobacterial isolates.

机译：用于检测产ESBL肠杆菌中质粒介导的喹诺酮耐药qnr基因的多重PCR。

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摘要

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 qnrB-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.
机译：目的：开发一种快速可靠的基于单管的PCR技术，以同时检测质粒介导的喹诺酮耐药性qnrA，qnrB和qnrS基因。方法：经过多次比对后，设计引物以检测已知的qnr变体（qnrA-为六个，qnrB-为六个，qnrS-like基因为两个）。它们被用来筛选从2002年至2004年收集的来自科威特的64种产生广谱β-内酰胺酶（ESBL）的肠杆菌分离株，因为ESBL基因通常与qnr基因相关。进行测序以鉴定qnr和相关的ESBL基因。结果：在优化的条件下，所有阳性对照（单独使用或混合使用）均证实了PCR引物的特异性。在64个分离株中，只有3个分离株的qnrB样基因呈阳性（4.7％），而未检测到qnrA样和qnrS样基因。在阴沟肠杆菌K34（SHV-12 +）分离物中检测到qnrB2基因，而从阴沟肠杆菌K37（SHV-12 +）和柠檬酸杆菌中鉴定出qnrB1样（称为qnrB7）和qnrB6样（称为qnrB8）基因。 freundii K70（VEB-1b +）分离株。结论：我们在这里报告了一种快速可靠的技术，用于快速筛查用于流行病学调查的qnr阳性菌株。在使用科威特分离株的研究中，ESBL产肠杆菌科细菌中Qnr决定簇的患病率较低。

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