MALDI-TOF-MS在鉴定进口甲鱼卵病原菌中的应用  

Application of MALDI-TOF-MS for Identifi cation of Pathogenic Bacteria in Imported Terrapin Eggs

作　　者：徐淑菲[1] 吴玉娟[2] 孔繁德[1] 赵冉 彭小莉[1] 叶妍妍[1] 陈信忠[1] 

机构地区：[1]厦门出入境检验检疫局,福建厦门361012 [2]集美大学生物工程学院,福建厦门361021 [3]厦门市农产品质量安全检验测试中心,福建厦门361000

出　　处：《中国动物检疫》2014年第7期86-90,100,共6页China Animal Health Inspection

基　　金：厦门市科技计划项目(项目编号:3502Z20124003)

摘　　要：目的应用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)仪器和Biotyper分析软件对台湾进口的甲鱼卵进行携带病原菌的检测。方法选用合适的培养基对进境甲鱼卵进行细菌分离培养,选取可疑菌株,纯化后接种肉汤培养,然后按照MALDI-TOF-MS要求进行样品制备,点靶并获取样品质谱图,并运用Biotyper分析软件对所获取的质谱图进行分析。同时用全自动微生物鉴定系统进行辅助分析鉴定。结果应用MALDITOF-MS方法从台湾输入大陆的73批甲鱼卵中分离出绿脓杆菌、奇异变形杆菌、肺炎克雷伯氏菌、霍乱弧菌、沙门氏菌等25个属49个种242株致病菌,其中大部分是对人和动物的条件致病菌。结论当前进境甲鱼卵携带病原菌形势严峻,需引起进出口商、检验机构及其他相关部门重视。本研究对规范甲鱼卵贸易、保障人民健康都有着重要的作用。Objective Matrix-Assisted Laser Desorption/ Ionization Time of Flight Mass Spectrometry（MALDI-TOFMS）instrument and the Biotyper analysis software was applied to investigate the pathogens carried in terrapin eggs from Taiwan. Methods Suitable media for the bacteria isolation and culture of the imported soft-shell turtle eggs were chosen,and suspicious bacterial strains were cultured,isolated and purifi ed,and then inoculated into broth. After being cultured in broth,the probable pathogens were prepared and added to the target polished steel plate and the mass spectra were acquired. The spectra were analyzed by the Biotyper analysis software. And automatic bacteria identifi cation system was used to aid the analysis and identifi cation of the pathogens. Results 242 bacteria strains in 49 species of 25 genera were isolated from the 73 lots of terrapin eggs from Taiwan by MALDI-TOF-MS,including Pseudomonas aeruginosa,Proteus mirabilis,Klebsiella pneumoniae,Salmonella,Vibrio cholera and most of them were opportunistic pathogens both for human and animals. Conclusion It was shown that the current situation of the imported terrapin eggs carrying pathogenic bacteria was serious,and the exporter or importer,the inspection organs and the other relevant departments are required to pay more attention to it. The study will play an important role in promoting the terrapin egg trade and protecting the public health.

关 键 词：甲鱼卵 MALDI-TOF-MS 质谱图 病原菌 

分 类 号：S851.347[农业科学—预防兽医学] S947.12[农业科学—兽医学]