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Improvement of MALDI-TOF MS profiling for the differentiation of species within the Acinetobacter calcoaceticus-Acinetobacter baumannii complex

机译:改进MALDI-TOF MS谱图以区分钙不动杆菌-鲍曼不动杆菌复合体中的物种

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MALDI-TOF MS is currently becoming the method of choice for rapid identification of bacterial species. in routine diagnostics. Yet, this method suffers from the inability to differentiate reliably between some closely related bacterial species including those of the Acinetobacter calcoaceticus-Acinetobacter baumannii (ACB) complex, namely A. baumannii and Acinetobacter nosocomialis. In the present study, We evaluated a protocol which was different from that used in the Bruker Daltonics identification system (MALDI BioTyper) to improve species identification using a taxonomically precisely defined set of 105 strains representing the four validly named species of the ACB complex. The novel protocol is based on the change in matrix composition from alpha-cyano-4-hydroxycinnamic acid (saturated solution in water:acetonitrile:trifluoroacetic acid, 47.5:50:2.5, v/v) to ferulic acid (12.5 mg ml(-1) solution in water:acetonitrile:formic acid 50:33:17, v/v), while the other steps of sample processing remain unchanged. Compared to the standard protocol, the novel one extended the range of detected compounds towards higher molecular weight, produced signals with better mass resolution, and allowed the detection of species-specific signals. As a result, differentiation of A. nosocomialis and A. baumannii strains by cluster analysis was improved and 13 A. nosocomialis strains, assigned erroneously or ambiguously by using the standard protocol, were correctly identified. (C) 2013 Elsevier GmbH. All rights reserved.
机译:MALDI-TOF MS当前正成为快速鉴定细菌种类的首选方法。在常规诊断中。然而,该方法由于不能可靠地区分一些紧密相关的细菌种类,包括钙不动杆菌-鲍曼不动杆菌(ACB)复合物即鲍曼不动杆菌和医院不动杆菌。在本研究中,我们评估了一种协议,该协议不同于在布鲁克·道尔顿公司识别系统(MALDI BioTyper)中使用的协议,该协议使用分类学上精确定义的105株菌株代表ACB复合体的四个有效命名物种,从而改善了物种鉴定。该新方案基于从α-氰基-4-羟基肉桂酸(水:乙腈:三氟乙酸的饱和溶液,47.5:50:2.5,v / v的饱和溶液)到阿魏酸(12.5 mg ml(- 1)在水:乙腈:甲酸50:33:17中的溶液,v / v),而样品处理的其他步骤保持不变。与标准方案相比,该新型方案将检测到的化合物的范围扩大到更高的分子量,产生了具有更好质量分辨率的信号,并允许检测物种特异性信号。结果,通过聚类分析改善了农杆菌和鲍曼不动杆菌菌株的区分,并且正确鉴定了使用标准方案错误或歧义分配的13个农杆菌。 (C)2013 Elsevier GmbH。版权所有。

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