School of Molecular Bioscience, The University of Sydney, NSW 2006, Australia.
J Antimicrob Chemother. 2014 Jan;69(1):77-80. doi: 10.1093/jac/dkt312. Epub 2013 Aug 6.
To explore the cause of third-generation cephalosporin resistance in Australian Acinetobacter baumannii isolates belonging to global clone 1 (GC1).
GC1 isolates from Australia were tested for resistance to ceftazidime and cefotaxime using disc diffusion and MICs. PCR was used to determine the context of ISAba1-ampC configurations and amplicons were sequenced. The level of transcripts was measured using quantitative real-time PCR. Multilocus sequence typing was performed.
All ceftazidime- and cefotaxime-resistant isolates carried an appropriately oriented ISAba1 adjacent to the ampC gene and ISAba1 increased ampC transcripts 8-12-fold. In three isolates, the ampC gene next to ISAba1 was not in the normal chromosomal position. Instead, ISAba1 was 7 bp upstream of an additional copy of ampC located in a 3155 bp duplicated segment of the chromosome that differs from the resident GC1 segment by 2.3% but is almost identical to the corresponding region in several non-GC1 draft genomes. The duplicated segment is bounded by directly oriented copies of ISAba1 and flanked by a 9 bp direct duplication. This 5.5 kb transposon, named Tn6168, is in the same position in the chromosome of the three Australian isolates and the GC1 isolate AB0057. Tn6168 was also detected in an unrelated A. baumannii strain, where it was in a different location. The central part of Tn6168 was probably acquired from a sequence type ST32 (Institut Pasteur scheme) A. baumannii strain.
The ISAba1-ampC configuration, which increases ampC expression, can be part of a composite transposon Tn6168.
探索属于全球克隆 1 (GC1)的澳大利亚鲍曼不动杆菌分离株第三代头孢菌素耐药的原因。
使用圆盘扩散和 MIC 法检测来自澳大利亚的 GC1 分离株对头孢他啶和头孢噻肟的耐药性。PCR 用于确定 ISAba1-ampC 构型的上下文,扩增子进行测序。使用定量实时 PCR 测量转录本的水平。进行多位点序列分型。
所有头孢他啶和头孢噻肟耐药的分离株均携带适当取向的 ISAba1 紧邻 ampC 基因,ISAba1 将 ampC 转录物增加 8-12 倍。在三个分离株中,紧邻 ISAba1 的 ampC 基因不在正常染色体位置。相反,ISAba1 在位于染色体上的额外 ampC 拷贝的上游 7 bp 处,该拷贝位于 3155 bp 重复片段中,与驻留 GC1 片段的差异为 2.3%,但与几个非 GC1 草图基因组的相应区域几乎相同。重复片段由直接取向的 ISAba1 拷贝和侧翼的 9 bp 直接重复序列限定。这个 5.5 kb 的转座子命名为 Tn6168,位于三个澳大利亚分离株和 GC1 分离株 AB0057 的染色体的相同位置。Tn6168 也在一个无关的鲍曼不动杆菌菌株中被检测到,它位于不同的位置。Tn6168 的中心部分可能来自序列类型 ST32(巴斯德研究所方案)鲍曼不动杆菌菌株。
增加 ampC 表达的 ISAba1-ampC 构型可以是复合转座子 Tn6168 的一部分。