Identification and characterisation of Klebsiella pneumoniae and Pseudomonas aeruginosa clinical isolates with atypical ß-lactam susceptibility profiles using Orbitrap liquid chromatography-tandem mass spectrometry
Identification and characterisation of Klebsiella pneumoniae and Pseudomonas aeruginosa clinical isolates with atypical ß-lactam susceptibility profiles using Orbitrap liquid chromatography-tandem mass spectrometry
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使用 Orbitrap 液相色谱-串联质谱法对具有非典型 β-内酰胺敏感性的肺炎克雷伯菌和铜绿假单胞菌临床分离株进行鉴定和表征
DOI：
10.1101/2022.02.27.482154
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发表时间：
2022
期刊：
影响因子：
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通讯作者：
Takebayashi Y
中科院分区：
文献类型：
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作者：
Takebayashi Y
通过分析细菌DNA或蛋白质直接预测抗菌药物敏感性的可能性引起了人们的极大兴趣。我们报告使用肺炎克雷伯菌、大肠埃希菌、铜绿假单胞菌和鲍曼不动杆菌转化子来确定β-内酰胺酶阳性临床分离株β-内酰胺类药物敏感性的基线预测规则。然后，我们部署了一种强大且可重复性的猎枪蛋白质组学方法，通过参考我们在培养细菌和培养阳性血液提取液中的基线预测规则，鉴定β内酰胺酶阳性并预测β内酰胺类药物敏感性。然后，蛋白质组学和全基因组测序使我们能够表征K。肺炎和P。铜绿假单胞菌与预期的β-内酰胺类药物敏感性谱不同，反复扩展我们的预测规则。蛋白质组学在全基因组测序产生的信息之外增加了相当大的价值，允许考虑基因表达，而不仅仅是基因的存在。具体地说，是墨水。在肺炎克雷伯菌中，我们确定了RandramR调节突变之间的关键差异，并比较了OmpK36天冬氨酸-苏氨酸或甘氨酸-天冬氨酸二肽孔蛋白插入对头孢吡肟和碳青霉烯类抗生素敏感性的影响。INP。在铜绿假单胞菌中，我们鉴定了mexRversusna突变的基因表达效应的差异，并将这些差异与β-内酰胺类对高产AmpC酶和/或产生金属-β-内酰胺酶的菌株的MIC的差异有关。
There is significant interest in the possibility of predicting antibacterial drug susceptibility directly though the analysis of bacterial DNA or protein. We report the use ofKlebsiella pneumoniae, Escherichia coli, Pseudomonas aeruginosaandAcinetobacter baumanniitransformants to define baseline predictive rules for the β- lactam susceptibility profiles of β-lactamase positive clinical isolates. We then deployed a robust and reproducible shotgun proteomics methodology to identify β-lactamase positivity and predict β-lactam susceptibility by reference to our baseline predictive rules both in cultured bacteria and in extracts of culture-positive blood. Proteomics and whole genome sequencing then allowed us to characteriseK. pneumoniaeandP. aeruginosaisolates that differed from the expected β-lactam susceptibility profile, iteratively expanding our predictive rules. Proteomics added considerable value over and above the information generated by whole genome sequencing, allowing for gene expression, not just gene presence to be considered. Specifically, inK. pneumoniae, we identified key differences betweenacrRandramRregulatory mutations and compared the effects of OmpK36 Aspartate-Threonine or Glycine-Aspartate dipeptide porin insertions on susceptibility to cefepime and carbapenems. InP. aeruginosa, we identified differences in the gene expression effects ofmexRversusnalCmutations and related these to differences in β-lactam MICs against isolates hyper-producing AmpC β-lactamase and or producing a metallo-β-lactamase.