MALDI-TOF-MS鉴定布鲁氏菌方法建立和评价  

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

作　　者：汤旭[1] 肖迪[2] 张慧芳[2] 陈经雕[3] 毛玲玲[4] 崔步云[2] 张建中[2] 姜海[2] 

机构地区：[1]北京市海淀区疾病预防控制中心,北京100094 [2]中国疾病预防控制中心传染病预防控制所,传染病预防控制围家重点实验室,感染性疾病协同诊治协同创新中心,北京102206 [3]广东省疾病预防控制中心,广州511430 [4]辽宁省疾病预防控制中心,沈阳110005

出　　处：《中国人兽共患病学报》2016年第9期772-778,共7页Chinese Journal of Zoonoses

基　　金：国家自然科学基金(No.81271900)~~

摘　　要：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。Fast,accurate and convenient detection and sensibility and specificity of the method was evaluated too. identification of Brucella, MALDI-TOF-MS was established. The 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.

关 键 词：布鲁氏菌 MALDI-TOF-MS 鉴定 

分 类 号：R378.5[医药卫生—病原生物学]