Classification and identification of enterococci: a comparative phenotypic, genotypic, and vibrational spectroscopic study
Classification and identification of enterococci: a comparative phenotypic, genotypic, and vibrational spectroscopic study
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DOI：
10.1128/jcm.39.5.1763-1770.2001
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发表时间：
2001-05-01
影响因子：
9.4
通讯作者：
Naumann, D
中科院分区：
文献类型：
--
作者：
Kirschner, C;Maquelin, K;Naumann, D
在物种水平上快速准确地鉴定肠球菌是临床微生物学的一项基本任务，因为这些细菌已经成为世界范围内医院感染的主要原因之一。振动光谱技术(红外[IR]和拉曼光谱)可以提供潜在的替代传统的分型方法，因为它们是快速、易于操作和经济的。我们提出了一项使用表型、基因分型和振动光谱技术对6个不同物种的18株肠球菌菌株进行分型的比较研究。通过傅立叶变换(FT)-IR光谱结合层次聚类分析对细菌进行分类，与API和MicroScan等自动表型系统获得的结果相比，某些菌株的分类结果存在差异。使用基因分型方法的进一步诊断评估--即物种特异性连接酶和糖肽抗性基因的聚合酶链式反应(仅限于鉴定四种肠球菌)和16S RNA测序(鉴定肠球菌的“金标准”)--证实了FT-IR分类获得的结果。这些结果后来被三个不同的实验室复制，使用共聚焦拉曼光谱、FT-IR衰减全反射光谱和FT-IR显微光谱，证明了该技术的分辨能力和重复性。结论：振动光谱技术在临床微生物学研究中具有作为常规方法的巨大潜力。
Rapid and accurate identification of enterococci at the species level is an essential task in clinical microbiology since these organisms have emerged as one of the leading causes of nosocomial infections worldwide. Vibrational spectroscopic techniques (infrared [IR] and Raman) could provide potential alternatives to conventional typing methods, because they are fast, easy to perform, and economical. We present a comparative study using phenotypic, genotypic, and vibrational spectroscopic techniques for typing a collection of 18 Enterococcus strains comprising six different species. Classification of the bacteria by Fourier transform (FT)-IR spectroscopy in combination with hierarchical cluster analysis revealed discrepancies for certain strains when compared with results obtained from automated phenotypic systems, such as API and MicroScan. Further diagnostic evaluation using genotypic methods-i.e., PCR of the species specific ligase and glycopeptide resistance genes, which is limited to the identification of only four Enterococcus species and 16S RNA sequencing, the "gold standard" for identification of enterococci-confirmed the results obtained by the FT-IR classification. These results were later reproduced by three different laboratories, using confocal Raman microspectroscopy, FT-IR attenuated total reflectance spectroscopy, and FT-IR microspectroscopy, demonstrating the discriminative capacity and the reproducibility of the technique. It is concluded that vibrational spectroscopic techniques have great potential as routine methods in clinical microbiology.