痰潘多拉菌鉴定方法分析  

The methodological comparison of Pandoraea sputorum identification

作　　者：田宏攀 肖潇 程小欢[1] 郭清莲[1] 方芳[1] 乐英棒 周俊英[1] 李一荣[1] Tian Hongpan;Xiao Xiao;Cheng Xiaohuan;Guo Qinglian;Fang Fang;Yue Yingbang;Zhou Junying;Li Yirong(Department of Clinical Laboratory,Zhongnan Hospital of Wuhan University,Wuhan 430071,China)

机构地区：[1]武汉大学中南医院检验科,武汉430071

出　　处：《中华检验医学杂志》2020年第7期739-744,共6页Chinese Journal of Laboratory Medicine

基　　金：国家科技重大专项(2017ZX10103005,2020ZX09201007)。

摘　　要：目的:评估痰潘多拉菌生化反应、基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)和基因测序的鉴定结果,为临床微生物鉴定痰潘多拉菌提供更佳方案。方法:方法性能评价。收集2018年7至10月武汉大学中南医院住院患者血培养中分离的10株痰潘多拉菌,对其培养特性、菌落形态和染色特征进行研究和记录。然后分别通过手工生化鉴定(API 20NE)、全自动微生物鉴定分析系统(梅里埃VITEK 2 Compact、BD Phoenix-100、赛默飞世尔ARIS 2X)、MALDI-TOF MS(梅里埃VITEK MS和布鲁克MALDI Biotyper)和16S核糖体核糖核酸(rRNA)基因测序对痰潘多拉菌进行鉴定,并评估各方法的鉴定性能。结果:痰潘多拉菌是一种动力阴性、氧化酶阳性、触酶阳性的非发酵革兰阴性杆菌。10株痰潘多拉菌的API 20NE鉴定结果为反硝化无色杆菌、粪产碱菌或少见嗜铜菌,种属鉴定准确率0;VITEK 2 Compact鉴定结果显示其中6株为潘多拉菌属,3株为其他种属细菌(洋葱伯克霍尔德菌群、少动鞘氨醇单胞菌、皮氏罗尔斯顿菌),1株为Low Discrim(低分辨率),属鉴定准确率6/10;Phoenix-100鉴定结果显示10株均为无鉴定结果,种属鉴定准确率0;ARIS 2X鉴定结果显示其中7株为其他种属细菌(嗜麦芽窄食单胞菌、鲁氏不动杆菌、少动鞘氨醇单胞菌、浅黄假单胞菌、鲍曼不动杆菌复合体/溶血不动杆菌),3株为极罕见生物类型,种属鉴定准确率0;VITEK MS和MALDI Biotyper鉴定结果均为痰潘多拉菌,种属鉴定准确率10/10;16S rRNA基因测序结果与痰潘多拉菌相似度为100%。结论:目前基于生化反应的临床微生物学检验方法很难将痰潘多拉菌准确鉴定到种的水平,MALDL-TOF MS和16S rRNA基因测序技术是目前鉴定痰潘多拉菌快速准确的方法。Objective The matria-assisted laser desorption/ionization time of flight mass spectrometry(MALDI-TOF MS)and sequencing methods were performed to assess the methodology(biochemistry methods)for identifying the Pandoraea sputorum and provide the more preferred approach to identify Pandoraea species.Methods This paper is a study on performance evaluation of identification method.Ten lines of Pandoraea sputorum were isolated from blood cultures of inpatients were collected at Zhongnan Hospital of Wuhan University from July to October 2018 and were confirmed by the cultural characteristics,colonial morphology and Gram′s stain.Further identification was carried out by using the manual biochemical method(API 20NE),automatic biochemistry systems(BioMerieuxVITEK 2 Compact,BD Phoenix-100andThemo ARIS 2X),MALDI-TOF MS(BioMerieuxVITEK MS and Bruker MALDI Biotyper)and the sequencing methods of the 16S rRNA to identify the Pandoraea sputorum.Results Pandoraea sputorum was non-fermented gram-negative bacteria that are non-motile,oxidase positive,and catalase positive.Ten lines of Pandoraea sputorum were identified as Achromobacter denitrificans,Alcaligenes faecalis or Cupriavidus pauculus and the accuracy rate of genus and species identification was 0 by API 20NE.Among all the samples,six lines were identified as the Pandoraea spp.with the accuracy rate of genus identification was 6/10 by VITEK 2 Compact;whereas the other four lines were identified as the Burkholderia cepacia,Sphingomonas paucimobilis,Ralstonia pickettii,or"Low Discrim".All of these were identified as"No Identification"by Phoenix-100,which the accuracy rate of genus and species identification was 0.Seven isolates were identified by ARIS 2X as Stenotrophomonas maltophilia,Acinetobacter lwoffii,Sphingomonas paucimobilis,Pseudomonas luteola,Acinetobacter baumannii/Acinetobacter haemolyticus;whereas the other three lines were identified as rare species,thus,the accuracy rate of genus and species identification was 0.Both VITEK MS and MALDI Biotyper indicated all

关 键 词：伯克霍尔德杆菌科 RNA 核糖体 16S 光谱法 质量 基质辅助激光解吸电离 痰 

分 类 号：R446.5[医药卫生—诊断学]