MALDI-TOF-MS鉴定布鲁氏菌方法建立和评价

目的 为了快速、准确、便捷的检测和鉴定布鲁氏菌,本研究拟建立鉴定布鲁氏菌属的MALDI-TOF-MS方法,利用现场分离菌株对该方法进行特异性和敏感性评价。方法 收集布鲁氏菌标准菌株和地方流行株,应用MALDI-TOF-MS采集图谱,获取独特的蛋白质指纹图谱,汇总成标准图谱,建立布鲁氏菌鉴定数据库。并用39株布鲁氏菌菌株,对建立的数据库进行验证。结果 经数据库鉴定,39株布鲁氏菌菌株与数据库中布鲁氏菌的匹配分数全部大于2.300,均报告为布鲁氏菌,表明鉴定结果的可信度很高。 聚类分型结果表明,在蛋白质水平,MALDI-TOF-MS将测试的布鲁氏菌分成3大类。结论 MALDI-TOF-MS对布鲁氏菌进行鉴定,具有快速、准确、灵敏等优点,可以实现对布鲁氏菌属的准确鉴定,对布鲁氏菌病的临床诊断具有较高的使用价值。

Establishment and evaluation of identification of Brucella by MALDI-TOF-MS

Fast,accurate and convenient detection and identification of Brucella,MALDI-TOF-MS was established. The sensibility and specificity of the method was evaluated too. MALDI-TOF-MS database was established by collecting the unique protein fingerprint of 20 Brucella reference strains and endemic strains and was evaluated by 39 Brucella isolates. On the basis of database information,59 Brucella strains were grouped into 3 clusters. Mass spectra database of Brucella identification was created through the acquisition of the Brucella characteristic fingerprint. Brucella isolates were selected to verify established database. Match scores between 39 strains and reference strains in database were all greater than 2.300 and results are reported as Brucella. This revealed that degree of confidence was very high. Clustering results indicated that Brucella was divided into three categories at the protein level by MALDI-TOF-MS. MALDI-TOF-MS was used to identify Brucella at protein levels. The method is fast,accurate,sensitive and has higher value on clinical diagnosis of brucellosis.