Rapid label-free identification of Klebsiella pneumoniae antibiotic resistant strains by the drop-coating deposition surface-enhanced Raman scattering method
Rapid label-free identification of Klebsiella pneumoniae antibiotic resistant strains by the drop-coating deposition surface-enhanced Raman scattering method
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DOI：
10.1016/j.saa.2017.04.044
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发表时间：
2017-08-05
影响因子：
4.4
通讯作者：
Lee, Hee Joo
中科院分区：
文献类型：
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作者：
Cheong, Youjin;Kim, Young Jin;Lee, Hee Joo
尽管已经开发了许多方法来鉴定未知细菌，但临床微生物学中的细菌鉴定仍然是一个复杂且耗时的过程。为了解决这个问题，我们开发了一种无标记的方法，用于快速鉴定临床相关的多位点测序分型验证的喹诺酮类耐药肺炎克雷伯菌菌株。我们还应用该方法鉴定了来自菌落样品的三个菌株，ATCC 70063（对照），ST 11和ST 15;这些是流行的喹诺酮类耐药K。pneumoniae菌株。使用滴涂沉积表面增强拉曼散射（DCD-SERS）程序结合多元统计方法鉴定菌落。我们的工作流程显示出拉曼强度的增强因子为11.3 × 10（6），重现性高（相对标准偏差为7.4%），检测灵敏度（100 pM罗丹明6 G），相关系数为0.98。所有喹诺酮类耐药克雷伯氏菌与大肠杆菌菌株相比，肺炎杆菌菌株显示出相似的光谱拉曼位移（高相关性），而与细菌类型无关，以及不同的拉曼振动模式。我们提出的DCD-SERS程序加上基于多元线性的鉴定方法在区分相似的微生物和肺炎克雷伯菌菌株的亚型方面取得了优异的性能。因此，我们的无标记DCD-SERS过程结合计算决策支持方法是一种潜在的有用的方法，用于快速鉴定临床相关的K。pneumoniae菌株(C)2017爱思唯尔B. V.保留所有权利。
Although many methodologies have been developed to identify unknown bacteria, bacterial identification in clinical microbiology remains a complex and time-consuming procedure. To address this problem, we developed a label-free method for rapidly identifying clinically relevant multilocus sequencing typing-verified quinolone-resistant Klebsiella pneumoniae strains. We also applied the method to identify three strains from colony samples, ATCC70063 (control), ST11 and ST15; these are the prevalent quinolone-resistant K. pneumoniae strains in East Asia. The colonies were identified using a drop-coating deposition surface-enhanced Raman scattering (DCD-SERS) procedure coupled with a multivariate statistical method. Our workflow exhibited an enhancement factor of 11.3 x 10(6) to Raman intensities, high reproducibility (relative standard deviation of 7.4%), and a sensitive limit of detection (100 pM rhodamine 6G), with a correlation coefficient of 0.98. All quinolone-resistant K. pneumoniae strains showed similar spectral Raman shifts (high correlations) regardless of bacterial type, as well as different Raman vibrational modes compared to Escherichia coli strains. Our proposed DCD-SERS procedure coupled with the multivariate statistics-based identification method achieved excellent performance in discriminating similar microbes from one another and also in subtyping of K pneumoniae strains. Therefore, our label-free DCD-SERS procedure coupled with the computational decision supporting method is a potentially useful method for the rapid identification of clinically relevant K. pneumoniae strains. (C) 2017 Elsevier B.V. All rights reserved.