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作 者:龚艳清[1] 郭书林[1] 陈信忠[1] 杨俊萍[1] 叶妍妍[1]
出 处:《中国动物检疫》2014年第6期89-93,共5页China Animal Health Inspection
基 金:国家质量监督检验检疫总局公益项目(201210055);国家质检总局科研项目(2013IK045)
摘 要:为建立快速检测和鉴定鱼类常见的爱德华氏菌的方法,采用基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF-MS)方法对来自淡水鱼的迟缓爱德华氏菌(Edwardsiella tarda)和鮰爱德华氏菌(E.ictaluri)进行检测和鉴定。为测试该方法的重复性和稳定性,采用营养肉汤、BHI琼脂、DHL和TSA等4种不同培养基,以及用肉汤分别培养24 h、36 h、48 h和72 h的菌株进行检测,结果表明对该菌的蛋白质谱图影响很小;冷冻保存2年的菌株,用MALDI-TOF-MS进行检测,鉴定分值变化很小。对分离自不同水生动物的迟缓爱德华氏菌菌株进行检测,均可得到准确的结果。表明MALDI-TOF-MS技术可以快速、准确地检测和鉴定这两种爱德华氏菌。Matrix assisted laser desorption/ionization time of flight mass spectrometry(MALDI-TOF-MS)was experimentally used to test and identify the reference strains Edwardsiella tarda and E. ictaluri isolated from freshwater fish to establish a rapid method for detection and identification of Edwardsiella sp. In order to verify its stability and repeatability,the bacteria cultured in different medium such as nutrient broth,BHI agar,DHL and TSA,and in different time intervals of 24,36,48 or 72 h were tested with the mass spectrometry and little impacts were found. There were no changes of the identification scores of the bacteria preserved for two years using MALDI- TOF-MS. The strains of E. tarda isolated from different aquatic animals could also be exactly identified by the method. It was shown that the MALDITOF-MS technique could be used to detect the two Edwardsiella sp. quickly and accurately.
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