优化差速离心法在血培养报警瓶MALDI-TOF MS细菌鉴定中的应用  

Application of the improved differential centrifugation method in the direct identification of bacteria from positive blood culture bottles by MALDI-TOF MS

作　　者：尚军[1] 李修远[1] 王丽赟[1] 汤燕燕[1] 李雪[1] 张韶娅 黄艳飞[1] 王玫[1] 张明新[1] 张桂[1] 孙宇峰[1] 顾海彤[1] 田小波[1] 鲁辛辛[1] 

机构地区：[1]首都医科大学附属北京同仁医院检验科,北京100730

出　　处：《临床检验杂志》2016年第12期913-918,共6页Chinese Journal of Clinical Laboratory Science

摘　　要：目的评价用于血培养报警瓶基质辅助激光解吸电离飞行时间质谱（matrix assisted laser desorptionionization-time of flight mass spectrometry,MALDI-TOF MS）细菌直接鉴定的前处理方法。方法建立优化差速离心法流程。收集首都医科大学附属北京同仁医院2015年10月至12月外周血培养报警标本100例（均为单一菌）,以转种培养后菌落MALDI-TOF MS鉴定为金标准,对比差速离心法优化前后、BRUKER SEPSITYPER试剂盒（简称试剂盒法）鉴定结果,评价优化差速离心法的准确率。收集2016年1月至10月外周血及体液培养报警标本516例（经革兰染色确认为细菌）,对优化差速离心法进行大样本应用研究。结果优化的差速离心法、试剂盒法及优化前的差速离心法鉴定准确率分别为97%、94%及92%,差异无统计学意义;优化的差速离心法鉴定分值〉2.0的例数则多于其他两种方法,与优化前相比差异具有统计学意义（χ~2=3.916,P=0.048）。516例样本单一菌486例,混合菌30例。优化的差速离心法鉴定准确率为92.2%（476/516）。外周血及其他体液样本鉴定准确率分别为96.5%（179/289）、86.8%（197/227）,差异有统计学意义（χ~2=16.92,P〈0.01）。单一菌鉴定准确率为97.1%（472/486）,其中革兰阳性球菌和阴性杆菌分别为96.9%、99.1%,均明显高于阳性杆菌（55.6%）,差异有统计学意义（χ~2=25.329,P〈0.01;χ~2=48.526,P〈0.01）。混合菌鉴定准确率为13.3%（4/30）。体液样本混合菌比例为11%（25/227）,显著高于外周血[1.7%（5/289）,χ~2=20.008,P〈0.01]。结论优化差速离心法鉴定率高、成本低、易操作,优于试剂盒法,对单一细菌MALDI-TOF MS血培养报警直接鉴定更具临床应用价值。Objective To evaluate a pretreatment method for the direct identification of bacteria from positive blood culture bottles by matrix-assisted laser desorption ionization-time of flight mass spectrometry（ MALDI-TOF MS）. Methods A modified differential centrifugation method was established. One hundred of positive blood culture samples with monomicrobic infection were collected from Beijing Tongren Hospital,Capital Medical University during October 2015 and December 2015. The identification results of traditional and improved differential centrifugation methods and the SEPSITYPER kit（ BRUKER,Germany） were evaluated,and compared with that of the MALDI-TOF MS as gold standard. In addition,516 of positive peripheral blood and body fluid samples with the confirmation of Gram＇s staining were collected from our hospital during January 2016 and October 2016,and used to verify the improved differential centrifugation method. Results The accuracy rates of the improved differential centrifugation method,the kit method and the traditional differential centrifugation method were 97%,94% and 92%,respectively,and there was no significant difference between them. The number of samples with above 2. 0 of identification score in the improved differential centrifugation method was more than that in the other two methods,and there was significant difference between the improved and traditional differential centrifugation method（ χ2= 3. 916,P = 0. 048）. Among 516 samples with positive Gram＇s staining,486 belonged to monomicrobic infection and the other30 to mixed infection. The accuracy rate of the improved differential centrifugation method was 92. 2%（ 476 /516）,and those from peripheral blood and the other body fluid samples were 96. 5%（ 179 /289） and 86. 8%（ 197 /227）,respectively,with significant difference between them（ χ2= 16. 92,P 〈0. 01）. The accuracy rate for monomicrobic infections was 97. 1%（ 472 /486）,and the accuracy rates of Gram-positive cocci and Gram-negative bacilli（ 96. 9% an

关 键 词：优化差速离心法 血培养瓶报警 基质辅助激光解吸电离飞行时间质谱 

分 类 号：R446.5[医药卫生—诊断学]