临床常见革兰氏阳性球菌蛋白指纹库的构建

为建立临床常见革兰氏阳性球菌的蛋白指纹库,为快速鉴定这些细菌奠定基础,收集了从临床中分离获得的185株革兰氏阳性球菌,包括金黄色葡萄球菌、表皮葡萄球菌、溶血性葡萄球菌、粪肠球菌和屎肠球菌。将这些菌株分成建模组和验证组,利用表面增强激光解析电离飞行时间质谱检测细菌蛋白,用ProteinChip和Biomarker Wizard软件对建模组细菌数据进行分析,筛选出每种细菌各自稳定表达的蛋白峰,并将数据导入自建的Fingerwave软件建立了临床常见革兰氏阳性球菌的蛋白指纹库。随后,将验证组细菌的蛋白峰数据与蛋白指纹库中蛋白峰数据进行相似度分析,以评价其鉴定符合率。建立了包含5种临床常见革兰氏阳性球菌的蛋白指纹库,利用其对验证组菌株进行鉴定,与应用传统微生物学鉴定及分子生物学方法获得的鉴定结果的符合率为100%。结果表明,进一步扩大并完善革兰氏阳性球菌的蛋白指纹库,将为临床病原菌的快速鉴定提供可能。

Establishment of Protein Fingerprint Database of Common Clinical Gram Positive Cocci

To establish a protein fingerprint database of common clinical Gram positive cocci, which can be applied for rapid identification of bacteria, 185 clinical bacterial isolates including Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus haemolyticus, Enterococcus faecium, Enterococcus faecalis were collected and studied. These isolates were assigned into training set and testing set. Bacterial proteins were detected by surface enhanced laser desorption &; ionization time-of-flight mass spectrometry (SELDI-TOF MS). Protein fingerprint was analyzed by ProteinChip and Biomarker Wizard software and a protein fingerprint database of some Gram positive cocci was established by using self-constructed Fingerwave software. Similarity analysis between the protein fingerprint of testing set and the constructed database was conducted to evaluate its diagnostic value. A small protein fingerprint database was developed for five common Gram positive cocci. All the tested strains using the database were successfully identified with a result perfectly consistent with those obtained by using automated microbiology analysis system and molecular biological method. Thus, construction of a large-scale protein fingerprint database is desired, which will provide a practicable tool for rapid identification of bacteria.