用表面增强激光解析／电离飞行时间质谱蛋白指纹快速鉴定4种临床常见菌

Rapid identification of four common bacteria by SELDI-TOF MS protein fingerprints

目的建立临床常见细菌蛋白指纹数据模型，为细菌的快速鉴定奠定基础。方法利用表面增强激光解析／电离飞行时间质谱检测大肠埃希菌、肺炎克雷伯菌、铜绿假单胞菌和金黄色葡萄球菌参考株的细菌蛋白，筛选每种细菌稳定表达的蛋白峰，建立细菌的蛋白指纹数据模型，将其数据导入自建Fingerwave软件。收集临床分离的大肠埃希菌、肺炎克雷伯菌、铜绿假单胞菌和金黄色葡萄球菌共256株并运用质谱仪检测其细菌蛋白峰，蛋白峰数据与Fingerwave软件中蛋白指纹数据模型进行相似度比较，以评价其鉴定符合率。结果初步建立了4种临床常见细菌的蛋白指纹数据模型，利用其对临床菌株进行鉴定，鉴定结果与传统微生物学鉴定及分子生物学鉴定结果的符合率分别为大肠埃希菌93．1％（54／58）、肺炎克雷伯菌87．2％（75／86）、铜绿假单胞菌95．2％（60／63）和金黄色葡萄球菌96．2％（51／53）。结论通过蛋白指纹数据的相似度比较，可同时对4种常见临床细菌进行鉴定，为细菌感染的快速诊断提供了可能性。

Objective To establish protein fingerprints of common bacteria in clinics and to lay a foundation for rapid identification of bacteria. Methods Strains of Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa and Staphylococcus aureus were detected by surface enhanced laser desorption and ionization time-of-flight mass spectrometry （SELDI-TOF MS）. Stable expression protein peaks were screened and the data was input into the self-constructed Fingerwave software for identification of target bacteria by protein fingerprint comparison. Two hundred and fifty-six clinical isolates, including E. coli, K. pneumoniae, P. aeruginosa and S. aureus were detected and the data was compared with constructed database to evaluate its diagnostic value. Results The protein fingerprints including four common bacteia was used to identify the target bacteria with identification rate of 93. 1% （54/58） for E. coli, 87.2% （75/86） for K. pneumoniae, 96.2% （60/63） for P. aeruginosa and 96.2% （51/53） for S. aureus, respectively. Con- clusion Common bacteria can be rapidly identified by using the protein fingerprint comparison, which pro- vides a powerful tool for bacterial identification.