Endimiani Andrea, Hujer Andrea M, Perez Federico, Bethel Christopher R, Hujer Kristine M, Kroeger Jennifer, Oethinger Margret, Paterson David L, Adams Mark D, Jacobs Michael R, Diekema Daniel J, Hall Gerri S, Jenkins Stephen G, Rice Louis B, Tenover Fred C, Bonomo Robert A
Louis Stokes Cleveland Department of Veterans Affairs Medical Center, Research Service, Cleveland, OH 44106, USA.
J Antimicrob Chemother. 2009 Mar;63(3):427-37. doi: 10.1093/jac/dkn547. Epub 2009 Jan 20.
The emergence of bla(KPC)-containing Klebsiella pneumoniae (KPC-Kp) isolates is attracting significant attention. Outbreaks in the Eastern USA have created serious treatment and infection control problems. A comparative multi-institutional analysis of these strains has not yet been performed.
We analysed 42 KPC-Kp recovered during 2006-07 from five institutions located in the Eastern USA. Antimicrobial susceptibility tests, analytical isoelectric focusing (aIEF), PCR and sequencing of bla genes, PFGE and rep-PCR were performed. Results By in vitro testing, KPC-Kp isolates were highly resistant to all non-carbapenem beta-lactams (MIC(90)s >or= 128 mg/L). Among carbapenems, MIC(50/90)s were 4/64 mg/L for imipenem and meropenem, 4/32 mg/L for doripenem and 8/128 for ertapenem. Combinations of clavulanate or tazobactam with a carbapenem or cefepime did not significantly lower the MIC values. Genetic analysis revealed that the isolates possessed the following bla genes: bla(KPC-2) (59.5%), bla(KPC-3) (40.5%), bla(TEM-1) (90.5%), bla(SHV-11) (95.2%) and bla(SHV-12) (50.0%). aIEF of crude beta-lactamase extracts from these strains supported our findings, showing beta-lactamases at pIs of 5.4, 7.6 and 8.2. The mean number of beta-lactamases was 3.5 (range 3-5). PFGE demonstrated that 32 (76.2%) isolates were clonally related (type A). Type A KPC-Kp isolates (20 bla(KPC-2) and 12 bla(KPC-3)) were detected in each of the five institutions. rep-PCR showed patterns consistent with PFGE.
We demonstrated the complex beta-lactamase background of KPC-Kp isolates that are emerging in multiple centres in the Eastern USA. The prevalence of a single dominant clone suggests that interstate transmission has occurred.
含bla(KPC)的肺炎克雷伯菌(KPC-Kp)分离株的出现引起了广泛关注。美国东部地区的疫情爆发带来了严重的治疗和感染控制问题。尚未对这些菌株进行多机构的比较分析。
我们分析了2006年至2007年期间从美国东部五个机构收集的42株KPC-Kp。进行了药敏试验、分析性等电聚焦(aIEF)、bla基因的PCR和测序、脉冲场凝胶电泳(PFGE)以及重复序列聚合酶链反应(rep-PCR)。结果 通过体外试验,KPC-Kp分离株对所有非碳青霉烯类β-内酰胺类药物高度耐药(MIC90s≥128mg/L)。在碳青霉烯类药物中,亚胺培南和美罗培南的MIC50/90s分别为4/64mg/L,多利培南为4/32mg/L,厄他培南为8/128mg/L。克拉维酸或他唑巴坦与碳青霉烯类或头孢吡肟联合使用并未显著降低MIC值。基因分析显示,这些分离株具有以下bla基因:bla(KPC-2)(59.5%)、bla(KPC-3)(40.5%)、bla(TEM-1)(90.5%)、bla(SHV-11)(95.2%)和bla(SHV-12)(50.0%)。对这些菌株的粗β-内酰胺酶提取物进行的aIEF支持了我们的发现,显示出pI值为5.4、7.6和8.2的β-内酰胺酶。β-内酰胺酶的平均数量为3.5(范围为3至5)。PFGE显示32株(76.2%)分离株具有克隆相关性(A类型)。在五个机构中的每个机构都检测到了A类型的KPC-Kp分离株(20株bla(KPC-2)和12株bla(KPC-3))。rep-PCR显示的模式与PFGE一致。
我们证明了在美国东部多个中心出现的KPC-Kp分离株具有复杂的β-内酰胺酶背景。单一优势克隆的流行表明已发生州际传播。