Modarresi Farzan, Azizi Omid, Shakibaie Mohammad Reza, Motamedifar Mohammad, Valibeigi Behnaz, Mansouri Shahla
Department of Microbiology and Virology, Afzalipour School of Medicine, Kerman, Iran.
Research Center for Infectious Diseases and Tropical Medicine, Kerman, Iran.
APMIS. 2015 Nov;123(11):959-68. doi: 10.1111/apm.12455. Epub 2015 Sep 9.
Resistance-nodulation-division efflux system (RND) adeABC contributes to intrinsic resistance to various drug classes in Acinetobacter baumannii. Similarly, quorum sensing (QS) plays an important role in the biofilm formation and pathogenicity of this bacterium. The aims of this study were to evaluate the influence of iron limitation on the expression of efflux pump (adeABC) genes and QS (luxI, luxR) system by relative quantitative real-time polymerase chain reaction (qRT-PCR). In addition, DNA sequence and phylogenetic relatedness of biofilm-associated protein (Bap) gene was also investigated. Sixty-five multidrug-resistant isolates of A. baumannii were recovered from ICU patients of three hospitals in Kerman, Iran. The isolates were highly resistant to at least 11 antibiotics (MIC ≥64 μg/mL); however, 87% and 89% were susceptible to colistin and tigecycline, respectively (MIC 0.05 μg/mL) (p ≤ 0.05). We detected the presence of RND efflux pump, QS, and bap genes with the frequencies of 92% (adeA), 61.5% (adeB), 84.6% (adeC), 80% (luxI), 61% (luxR), and 66% (bap), respectively. qRT-PCR analysis showed that in some isolates, expression of both adeABC and luxI/R was increased more than fourfold in the presence of low iron (20 μm), suggesting the additional regulatory role of iron on both efflux pump and QS system. Alignment and phylogenetic analysis on the strong biofilm forming isolates confirmed that the fragments amplified were indeed part of bap gene and deduced sequence was similar to A. baumannii K9B410.
耐药-结瘤-分裂外排系统(RND)adeABC有助于鲍曼不动杆菌对多种药物类别的固有耐药性。同样,群体感应(QS)在该细菌的生物膜形成和致病性中起重要作用。本研究的目的是通过相对定量实时聚合酶链反应(qRT-PCR)评估铁限制对外排泵(adeABC)基因和QS(luxI、luxR)系统表达的影响。此外,还研究了生物膜相关蛋白(Bap)基因的DNA序列和系统发育相关性。从伊朗克尔曼三家医院的重症监护病房患者中分离出65株多重耐药鲍曼不动杆菌。这些分离株对至少11种抗生素高度耐药(MIC≥64μg/mL);然而,分别有87%和89%对黏菌素和替加环素敏感(MIC 0.05μg/mL)(p≤0.05)。我们检测到RND外排泵、QS和bap基因的存在频率分别为92%(adeA)、61.5%(adeB)、84.6%(adeC)、80%(luxI)、61%(luxR)和66%(bap)。qRT-PCR分析表明,在一些分离株中,在低铁(20μm)存在的情况下,adeABC和luxI/R的表达均增加了四倍以上,表明铁对外排泵和QS系统均有额外的调节作用。对强生物膜形成分离株的比对和系统发育分析证实,扩增的片段确实是bap基因的一部分,推导的序列与鲍曼不动杆菌K9B410相似。