基于拉曼光谱的成团泛菌快速鉴别  

Rapid identification of Pantoea agglomerans based on Raman spectroscopy

作　　者：宋莉 张孟 李悦 郝泽壮 丁长虹[3] 赵妍 周长龙[3] 段明华 Song Li;Zhang Meng;Li Yue;Hao Zezhuang;Ding Changhong;Zhao Yan;Zhou Changlong;Duan Minghua(Microbiological Testing Department,Changchun Center for Food and Drug Control,Changchun,Jilin,130000,China;Changchun University of Chinese Medicine,Changchun,Jilin,130117,China;Aviation University Air Force,Changchun,Jilin,130000,China;The 4th Air Force Station 0f PLA Air Force Harbin Flight Academy,Jinzhou,121000,China.)

机构地区：[1]长春市食品药品检验中心微生物检验科室,长春130000 [2]长春中医药大学 [3]空军航空大学 [4]哈尔滨飞行学院第四训练旅锦州场站卫生队

出　　处：《西南国防医药》2021年第4期314-318,共5页Medical Journal of National Defending Forces in Southwest China

摘　　要：目的对药品中存在的成团泛菌进行纯化与鉴定,建立应用共聚焦显微拉曼光谱仪快速准确判定药品微生物限度成团泛菌的方法。方法药品进行微生物限度检测,对需要检测的微生物范围之外的可疑菌落,对可疑菌落进行培养,分离纯化。然后进行种属鉴别,蛋白质鉴别和拉曼光谱鉴别。结果在木糖赖氨酸脱氧胆酸钠培养基(xylose lysine desoxycholate medium,XLD)选择性培养基上获得了黄色黏性的菌落,经BD Phoenix M50细菌鉴定药敏分析仪对菌落进行鉴别,结果为成团泛菌,可信值为99.0%;经微生物鉴定飞行时间质谱仪鉴定菌落为成团泛菌,分值为1.97,鉴定结果可信;应用共聚焦显微拉曼色谱仪对成团泛菌进行了扫描,在970 cm^(-1)和1010 cm^(-1)处有明显拉曼特征峰。结论从可疑菌落的种属、含有的蛋白质和表面物质的拉曼特征吸收峰3个角度对菌落进行鉴别和定性,最终将可疑菌落判定为成团泛菌。该实验获得了成团泛菌拉曼特征峰,从而对药品中存在的微生物进行科学判定,准确反映药品的微生物限度,确保药品的安全性。Objective To purify and identify the Pantoea agglomerans in drugs,and to establish a rapid and accurate method to determine the microbial limit of Pantoea agglomerans in drugs by using a confocal microscopy Raman spectrometer.Methods Microbial limit test was carried out for drugs,and suspicious colonies other than the microorganisms in need of test were cultured,separated and purified.Then species identification,protein identification and Raman spectrum identification were carried out.Results Yellow viscous colonies were obtained on xylose lysine desoxycholate(XLD) selective medium.The colonies were identified by BD Phoenix M50 bacterial identification and drug sensitivity analyzer.The results showed that the colonies were Pantoea agglomerans with a confidence value of 99.0%.The colony was identified by microbial identification time-of-flight mass spectrometer(TOFMS),and the score was 1.97,indicating that the identification result was reliable.Pantoea agglomerans were scanned by a confocal microscopy Raman spectrometer,and there were obvious Raman characteristic peaks at 970 cm^(-1) and 1010 cm^(-1).Conclusion Suspicious colonies were identified and characterized from the Raman characteristic absorption peaks of the species,the protein and the surface substances,and finally the suspicious colonies were determined as Pantoea agglomerans.The Raman characteristic peaks obtained in this experiment enables to scientifically determine the microbes in the drug,accurately reflect the microbial limit of the drug,and ensure the safety of the drug.

关 键 词：共聚焦显微拉曼光谱仪 成团泛菌拉曼光谱 快速鉴别 

分 类 号：R969.3[医药卫生—药理学]