Prediction of Fluoroquinolone Susceptibility Directly from Whole-Genome Sequence Data by Using Liquid Chromatography-Tandem Mass Spectrometry To Identify Mutant Genotypes
Prediction of Fluoroquinolone Susceptibility Directly from Whole-Genome Sequence Data by Using Liquid Chromatography-Tandem Mass Spectrometry To Identify Mutant Genotypes
复制标题
利用液相色谱-串联质谱直接从全基因组序列数据预测对氟喹诺酮类药物的易感性以识别突变基因型
DOI：
10.1128/aac.01814-17
复制
发表时间：
2018-03-01
影响因子：
4.9
通讯作者：
Avison, Matthew B.
中科院分区：
文献类型：
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作者：
Ismah, Wan Ahmad Kamil Wan Nur;Takebayashi, Yuiko;Avison, Matthew B.
革兰氏阴性细菌中的氟喹诺酮耐药性是多因素的，包括靶位点突变、孔蛋白产生减少导致氟喹诺酮进入减少、氟喹诺酮流出增加、修饰氟喹诺酮的酶以及 Qnr（一种保护药物靶点免受氟喹诺酮结合的 DNA 模拟物）。在这里，我们使用肺炎克雷伯菌作为模型系统，利用转化和体外突变体选择，对氟喹诺酮不敏感性的每种机制的相对重要性进行了全面分析。我们改进的生物学理解随后被用来生成 47 条规则，可以预测肺炎克雷伯菌临床分离株的氟喹诺酮敏感性。这一预测过程成功的关键是使用液相色谱-串联质谱法来测量培养细菌提取物中的蛋白质丰度，识别全基因组序列数据中看到的哪些序列变异在氟喹诺酮敏感性背景下具有重要的功能。
Fluoroquinolone resistance in Gram-negative bacteria is multifactorial, involving target site mutations, reductions in fluoroquinolone entry due to reduced porin production, increased fluoroquinolone efflux, enzymes that modify fluoro-quinolones, and Qnr, a DNA mimic that protects the drug target from fluoroquinolone binding. Here we report a comprehensive analysis, using transformation and in vitro mutant selection, of the relative importance of each of these mechanisms for fluoroquinolone nonsusceptibility using Klebsiella pneumoniae as a model system. Our improved biological understanding was then used to generate 47 rules that can predict fluoroquinolone susceptibility in K. pneumoniae clinical isolates. Key to the success of this predictive process was the use of liquid chromatography-tandem mass spectrometry to measure the abundance of proteins in extracts of cultured bacteria, identifying which sequence variants seen in the whole-genome sequence data were functionally important in the context of fluoroquinolone susceptibility.