利用SELDI-TOF-MS快速鉴定鲍曼不动杆菌的研究

Rapid identification of Acinetobacter baumannii by using surface enhanced laser desorption/ionization time-of-flight mass spectrometry

目的利用表面增强激光解析电离飞行时间质谱(SELDI-TOF-MS)技术快速鉴定鲍曼不动杆菌。方法收集临床分离鉴定的鲍曼不动杆菌60株及对照菌160株,将其所有菌株分成训练组和验证组。运用SELDI-TOF-MS技术检测鲍曼不动杆菌和对照菌的蛋白表达,筛选鲍曼不动杆菌特征性蛋白。用Ciphergen Proteinchip软件自动采集数据,结果经BioMarker Wizard和Bio Marker Pattern分析建立分类树模型。利用该模型对20株鲍曼不动杆菌和51株非鲍曼不动杆菌进行盲法验证。结果在相对分子量3000～30000 Da范围内共检测到72个蛋白峰,其中具有统计学差异的蛋白峰有56个(P <0. 01)。Bio Marker Pattern软件判别分析,择优选出质荷比(M/Z)为1 6737. 8和5763. 61的蛋白质峰建立鲍曼不动杆菌分类树模型。盲法验证结果表明SELDI-TOF-MS对鲍曼不动杆菌诊断的灵敏性和特异性分别为85%(17/20)、100%(51/51)。结论利用SELDI-TOF-MS可快速鉴定鲍曼不动杆菌。

Objective To establish the method of surface enhanced laser desorption/ionization time-of-flight mass spectrometry( SELDI-TOF-MS) to rapidly identify Acinetobacter baumannii( A. baumannii). Methods A total of 60 A. baumannii strains and 160 control strains were collected.The strains were divided into training and testing groups.SELDI-TOF-MS was used to detect the protein profiling of the bacteria.Data were automatically collected by Ciphergen Proteinchip Software and protein markers of A.baumannii were screened by Bio Marker Wizard Software.Classification tree model was developed and validated by Bio Marker Patterns Software.After that,the model was blindly verified with twenty A.baumannii and fifty-one control bacteria.Results Seventy two peaks were detected between 3000 and 30000 Da,among which fifty-six peaks showed significantly difference between A. baumannii and the control bacteria( P < 0. 01).By using Bio Marker Patterns Software,proteins( m/z at 1 6737. 8 and 5763. 61) were chosen to develop a classification tree model.The sensitivity and specificity of the method were 85% and 100%,respectively.Conclusion SELDI-TOF-MS has the potential for rapid identification of A.baumannii.