a German Armed Forces Hospital of Hamburg, Department of Tropical Medicine at the Bernhard Nocht Institute , Hamburg , Germany.
b Department of Medical Microbiology, University Medical Center Göttingen , Göttingen , Germany.
Crit Rev Microbiol. 2017 May;43(3):263-293. doi: 10.3109/1040841X.2016.1169990. Epub 2017 Jan 27.
Early identification of microbial pathogens is essential for rational and conservative antibiotic use especially in the case of known regional resistance patterns. Here, we describe fluorescence in situ hybridization (FISH) as one of the rapid methods for easy identification of microbial pathogens, and its advantages and disadvantages for the diagnosis of pathogens in human infections in the laboratory diagnostic routine. Binding of short fluorescence-labeled DNA or nucleic acid-mimicking PNA probes to ribosomes of infectious agents with consecutive analysis by fluorescence microscopy allows identification of bacterial and eukaryotic pathogens at genus or species level. FISH analysis leads to immediate differentiation of infectious agents without delay due to the need for microbial culture. As a microscopic technique, FISH has the unique potential to provide information about spatial resolution, morphology and identification of key pathogens in mixed species samples. On-going automation and commercialization of the FISH procedure has led to significant shortening of the time-to-result and increased test reliability. FISH is a useful tool for the rapid initial identification of microbial pathogens, even from primary materials. Among the rapidly developing alternative techniques, FISH serves as a bridging technology between microscopy, microbial culture, biochemical identification and molecular diagnostic procedures.
早期识别微生物病原体对于合理和保守地使用抗生素至关重要,特别是在已知区域耐药模式的情况下。在这里,我们将荧光原位杂交(FISH)描述为一种快速方法,用于在实验室诊断常规中轻松识别人类感染中的微生物病原体,及其用于病原体诊断的优缺点。用荧光标记的短 DNA 或类似核酸的 PNA 探针与核糖体结合,然后通过荧光显微镜进行连续分析,可在属或种水平上鉴定细菌和真核病原体。由于不需要进行微生物培养,FISH 分析可立即进行鉴定,从而避免了延迟。作为一种微观技术,FISH 具有独特的潜力,可以提供有关混合物种样本中关键病原体的空间分辨率、形态和鉴定的信息。FISH 程序的持续自动化和商业化已经显著缩短了检测时间,并提高了检测可靠性。FISH 是快速初始鉴定微生物病原体的有用工具,即使是从原始材料中。在快速发展的替代技术中,FISH 是显微镜、微生物培养、生化鉴定和分子诊断程序之间的桥梁技术。