基质辅助激光解吸电离飞行时间质谱快速检测无菌体液分离病原菌的研究  

Study of matrix-assisted laser desorption ionization-time of flight mass spectrometry(MALDI-TOF MS)for rapid detection of pathogenic bacteria isolated from sterile body fluids

作　　者：王洁[1] 贾乾 李静[1] 杨朝菊[1] 帖彦清[1] WANG Jie;JIA Qian;LI Jing;YANG Chao-ju;TIE Yan-qing(Department of Clinical Laboratory,Hebei General Hospital,Shijiazhuang 050051,China;Medical Examination Center,Hebei General Hospital,Shijiazhuang 050051,China)

机构地区：[1]河北省人民医院检验科,河北石家庄050051 [2]河北省人民医院体检中心,河北石家庄050051

出　　处：《河北医科大学学报》2024年第9期1090-1096,共7页Journal of Hebei Medical University

基　　金：河北省医学科学研究课题计划(20220929)。

摘　　要：目的探讨基质辅助激光解吸电离飞行时间质谱(matrix-assisted laser desorption ionization-time of flight mass spectrometry,MALDI-TOF MS)快速检测增菌报阳后无菌液体病原菌的临床应用价值。方法收集河北省人民医院肉汤增菌培养阳性的无菌体液标本412份。报阳后的标本进行涂片革兰染色镜检,同时接种固体培养基进行培养,35℃培养4~6 h长出菌膜,35℃培养18~24 h长出单个菌落,应用质谱仪VITEK MS分别对菌膜和菌落进行鉴定,以质谱鉴定菌落的结果作为参考标准,分析菌膜的鉴定符合率。结果VITEK MS(菌落)共鉴定出412株病原菌,包括革兰阳性球菌212株(51.45%)、革兰阴性杆菌172株(41.75%)和真菌28株(6.80%)。与VITEK MS(菌落)鉴定结果相比,VITEK MS(菌膜)总体鉴定符合率为91.50%(377/412),革兰阳性球菌鉴定符合率为89.15%(189/212),革兰阴性杆菌鉴定符合率为94.19%(162/172),差异均有统计学意义(χ^(2)=36.553、24.319、10.299,均P<0.05);真菌鉴定,VITEK MS(菌膜)鉴定符合率为92.86%(26/28),差异无统计学意义(χ^(2)=2.074,P=0.150)。结论应用基质辅助激光解吸电离飞行时间质谱技术对报阳后的无菌体液标本进行4~6 h菌膜鉴定,可进一步缩短病原菌鉴定时间,快速准确提供鉴定结果,为临床侵袭性感染疾病的诊疗提供早期依据。Objective To explore the clinical application value of matrix-assisted laser desorption ionization-time of flight mass spectrometry(MALDI-TOF MS)for rapid detection of pathogenic bacteria isolated from sterile body fluids after positive reporting of enrichment.Methods Four hundred and twelve specimens of sterile body fluids with positive broth culture were collected from Hebei General Hospital.The positive specimens were microscopically examined by smear Gram staining,and inoculated with solid medium for culture.The bacterial membrane grew at 35℃for 4-6 h,and the single colony grew at 35℃for 18-24 h.The VITEK MS was applied to identify the bacterial membrane and the colony respectively,and the results of the mass spectrometry identification of the colony were used as the reference standard to analyze the identification rate of the mass spectrometry identification of the bacterial membrane.Results A total of 412 pathogenic strains were identified by VITEK MS(colony),including 212 Gram-positive cocci(51.45%),172 Gram-negative bacilli(41.75%)and 28 fungal strains(6.80%).Compared with the VITEK MS(colony)identification results,the overall identification compliance rate was 91.50%(377/412)for VITEK MS(membrane),89.15%(189/212)for Gram-positive cocci and 94.19%(162/172)for Gram-negative bacilli,all with significant differences(χ^(2)=36.553,24.319,10.299,P<0.05).For fungal identification,identification compliance rate was 92.86%(26/28)by VITEK MS(membrane),with no significant difference(χ^(2)=2.074,P=0.150).Conclusion The application of MALDI-TOF MS for 4-6 h identification of sterile body fluid specimens after positive reporting can further shorten the identification time of pathogenic bacteria,provide rapid and accurate identification results,and provide an early basis for the diagnosis and treatment of invasive infectious diseases in clinical practice.

关 键 词：细菌 质谱 无菌体液 快速鉴定 

分 类 号：R378[医药卫生—病原生物学]