Harz M, Rösch P, Popp J
Institute of Physical Chemistry, Friedrich-Schiller-Universität Jena, Helmholtzweg 4, Jena 07743, Germany.
Cytometry A. 2009 Feb;75(2):104-13. doi: 10.1002/cyto.a.20682.
Rapid microbial detection and identification with a high grade of sensitivity and selectivity is a great and challenging issue in many fields, primarily in clinical diagnosis, pharmaceutical, or food processing technology. The tedious and time-consuming processes of current microbiological approaches call for faster ideally on-line identification techniques. The vibrational spectroscopic techniques IR absorption and Raman spectroscopy are noninvasive methods yielding molecular fingerprint information; thus, allowing for a fast and reliable analysis of complex biological systems such as bacterial or yeast cells. In this short review, we discuss recent vibrational spectroscopic advances in microbial identification of yeast and bacterial cells for bulk environment and single-cell analysis. IR absorption spectroscopy enables a bulk analysis whereas micro-Raman-spectroscopy with excitation in the near infrared or visible range has the potential for the analysis of single bacterial and yeast cells. The inherently weak Raman signal can be increased up to several orders of magnitude by applying Raman signal enhancement methods such as UV-resonance Raman spectroscopy with excitation in the deep UV region, surface enhanced Raman scattering, or tip-enhanced Raman scattering.
在许多领域,尤其是临床诊断、制药或食品加工技术领域,快速且具有高灵敏度和选择性的微生物检测与鉴定是一个重大且具有挑战性的问题。当前微生物学方法繁琐且耗时,因此需要更快速、理想情况下是在线的鉴定技术。振动光谱技术(红外吸收和拉曼光谱)是产生分子指纹信息的非侵入性方法,从而能够对诸如细菌或酵母细胞等复杂生物系统进行快速可靠的分析。在这篇简短的综述中,我们讨论了用于大量环境和单细胞分析的酵母和细菌细胞微生物鉴定中振动光谱技术的最新进展。红外吸收光谱可进行大量分析,而在近红外或可见光范围内激发的显微拉曼光谱有潜力用于单个细菌和酵母细胞的分析。通过应用拉曼信号增强方法,如在深紫外区域激发的紫外共振拉曼光谱、表面增强拉曼散射或针尖增强拉曼散射,固有较弱的拉曼信号可增强几个数量级。