基质辅助激光解析电离-飞行时间质谱法对铜绿假单胞菌的鉴定与聚类分析

Identification and cluster analysis of Pseudomonas aeruginosa by matrix assisted laser desorption lonization-time of flight mass spectrometery

中文关键词:  铜绿假单胞菌  基质辅助激光解析电离-飞行时间质谱法  鉴定  聚类分析

英文关键词:Pseudomonas aeruginosa  matrix assisted laser desorption lonization-time of flight mass spectrometry  identification  cluster analysis

中文摘要:

      目的 建立基质辅助激光解析电离-飞行时间质谱法(matrix assisted laser desorption lonization-time of flight mass spectrometery, MALDI-TOF MS)快速鉴定和聚类分析铜绿假单胞菌(Pseudomonas aeruginosa, P. aeruginosa)。方法 将全自动微生物分析仪鉴定为P. aeruginosa的20株分离株和1株标准菌株(CICC 21636)进行MALDI-TOF MS检测。经离线微生物鉴定软件分析, 所有菌株均报告为P. aeruginosa。获取21株 P. aeruginosa的特征蛋白质指纹图谱, 建立命名为Pseudomonas aeruginosa的自建数据库, 并采用 P. aeruginosa标准菌株CICC 21636进行验证。结果 Pseudomonas aeruginosa自建数据库对P. aeruginosa的鉴定结果较设备自带数据库明显提高。而且自建数据库信息基础上, 进一步对21株P. aeruginosa进行聚类分型, 实现了对不同来源P. aeruginosa可能污染源的追溯。结论 作为一种快速、准确、高通量的全新技术手段, MALDI-TOF MS能够实现对P. aeruginosa的特异性快速鉴定, 能满足在公共安全卫生、突发食品安全事件和口岸快速通关方面对P. aeruginosa的快速鉴定需求。

英文摘要:

      Objective To establish a method for rapid identification and cluster analysis of Pseudomonas aeruginosa by matrix assisted laser desorption lonization-time of flight mass spectrometry (MALDI-TOF MS). Methods Totally 20 field isolates, which were identified as P. aeruginosa by automatic microbiology analyzer, and 1 standard strain (CICC 21636) were detected by MALDI-TOF MS. All strains were identified as P. aeruginosa by analysis of off-line microbial identification software. The characteristic protein fingerprints of 21 P. aeruginosa strains were used to establish a self-built database named Pseudomonas aeruginosa. The standard strain CICC 21636 of P. aeruginosa was used to verify the results. Results The identification results of P. aeruginosa using Pseudomonas aeruginosa self-built database were significantly higher than those using the equipment-owned database. Moreover, on the basis of self-built database information, 21 P. aeruginosa strains were further clustered and the dendrogram was made. Therefore, the traceability of possible pollution sources of P. aeruginosa from different sources was achieved. Conclusion As a fast, accurate and high throughput identification technology, MALDI-TOF MS could realize the specific identification of P. aeruginosa, and meet the requirement of rapid identification of P. aeruginosa in the aspects of the public safety and health, emergency food safety incidents and rapid customs clearance at ports.