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首页> 外文期刊>Journal of Clinical Microbiology >Evaluation of Fluorescence-Based Amplified Fragment Length Polymorphism Analysis for Molecular Typing in Hospital Epidemiology: Comparison with Pulsed-Field Gel Electrophoresis for Typing Strains of Vancomycin-Resistant Enterococcus faecium
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Evaluation of Fluorescence-Based Amplified Fragment Length Polymorphism Analysis for Molecular Typing in Hospital Epidemiology: Comparison with Pulsed-Field Gel Electrophoresis for Typing Strains of Vancomycin-Resistant Enterococcus faecium

机译：基于荧光的扩增片段长度多态性分析在医院流行病学中分子分型的评价：与脉冲场凝胶电泳分型耐万古霉素肠球菌菌株的比较

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Fluorescence-based amplified fragment length polymorphism (fbAFLP) is a novel assay based on the fluorescent analysis of an amplified subset of restriction fragments. The fbAFLP assay involves the selective PCR amplification of restriction fragments from a total digest of genomic DNA. The ligation of adapters with primer-specific sites coupled with primers containing selective nucleotides allowed the full potential of PCR to be realized while maintaining the advantages of restriction endonuclease analysis. Fluorescence-based fragment analysis with polyacrylamide gel electrophoresis provides the accurate band sizing required for homology assessment. The large number of phylogenetically informative characters obtained by fbAFLP is well suited for cluster analysis and database development. The method demonstrated excellent reproducibility and ease of performance and interpretation. We typed 30 epidemiologically well-characterized isolates of vancomycin-resistant enterococci from an outbreak in a university hospital by fbAFLP. Clustering of fbAFLP data matched epidemiological, microbiological, and pulsed-field gel electrophoresis data. This study demonstrates the unprecedented utility of fbAFLP for epidemiological investigation. Future developments in standardization and automation will set fbAFLP as the “gold standard” for molecular typing in epidemiology.
机译：基于荧光的扩增片段长度多态性（fbAFLP）是一种基于限制性片段的扩增子集的荧光分析的新型检测方法。 fbAFLP分析涉及从基因组DNA的总消化物中限制性片段的选择性PCR扩增。衔接子与引物特异性位点的连接，再加上含有选择性核苷酸的引物，使得在保持限制性核酸内切酶分析优势的同时，可以充分发挥PCR的潜力。聚丙烯酰胺凝胶电泳基于荧光的片段分析可提供同源性评估所需的准确条带大小。通过fbAFLP获得的大量系统信息学特征非常适合于聚类分析和数据库开发。该方法显示出极好的可重复性，并且易于操作和解释。我们通过fbAFLP在大学医院的一次暴发中输入了30种流行病学特征良好的耐万古霉素肠球菌分离株。 fbAFLP数据的聚类与流行病学，微生物学和脉冲场凝胶电泳数据匹配。这项研究表明fbAFLP在流行病学研究中具有空前的实用性。标准化和自动化的未来发展将使fbAFLP成为流行病学分子分型的“金标准”。

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