基于MALDI-TOF MS对脓肿分枝杆菌脓肿亚种和马赛亚种的鉴别及特征分析  

Identification of Mycobacterium abscessus subsp.abscessus and subsp.massiliense based on MALDI-TOF MS and analysis for their characteristics

作　　者：秦雪娅 林勇 周聪 张慧 徐茂锁 QIN Xueya;LIN Yong;ZHOU Cong;ZHANG Hui;XU Maosuo(Department of Laboratory Medicine,Shanghai Fifth People's Hospital,Fudan University,Shanghai 200240,China)

机构地区：[1]复旦大学附属上海市第五人民医院检验科,上海200240

出　　处：《临床检验杂志》2025年第2期81-87,共7页Chinese Journal of Clinical Laboratory Science

基　　金：上海市闵行区卫生健康系统优秀青年医技人才和药学人才培养项目计划(MWYJYX07);上海市闵行区医学特色专科建设项目(2025MWTZB01)。

摘　　要：目的基于基质辅助激光解吸电离飞行时间质谱(MALDI-TOFMS)对脓肿分枝杆菌进行亚种水平鉴定和特征分析。方法使用Biotyper软件构建脓肿分枝杆菌脓肿亚种和马赛亚种的预测峰谱,实验组和对照组分别以预期最大峰值数70和峰值频率100%构建预测峰谱,对31株未用于构建预测峰谱的脓肿分枝杆菌进行盲测,以评估预测峰谱的鉴定效能;使用FlexAnalysis软件对脓肿分枝杆菌质谱峰值列表汇总分析,筛选脓肿分枝杆菌不同亚种质谱的特异峰;采用主成分分析(PCA)算法对脓肿分枝杆菌质谱数据进行聚类分析,探索PCA聚类对脓肿分枝杆菌亚种区分的可行性。结果实验组96.8%(30/31)的菌株被正确鉴定,1株粗糙菌落形态的脓肿分枝杆菌马赛亚种被误识别为脓肿分枝杆菌脓肿亚种;对照组77.4%(24/31)的菌株被正确鉴定,7株脓肿分枝杆菌马赛亚种被错误鉴定或无法识别,实验组鉴定效能显著优于对照组,差异有统计学意义(χ^(2)=5.167,P=0.026);脓肿分枝杆菌脓肿亚种具有3个特异峰(m/z4001.67、4386.81和4963.17),脓肿分枝杆菌马赛亚种也具有3个特异峰(m/z4950.48、4381.78和5214.90);PCA三维散点图中脓肿分枝杆菌脓肿亚种和马赛亚种数据点相对分散,未出现明显聚集簇;PCA树状图分为6个分支,仅4个分支为单一亚种组成,脓肿分枝杆菌脓肿亚种和马赛亚种最小距离水平值约为0.1。结论基于MALDI-TOFMS以预期最大峰值数70的方式构建预测峰谱可在亚种水平上准确鉴定脓肿分枝杆菌;使用质谱特异峰区分脓肿分枝杆菌脓肿亚种和马赛亚种具有可行性,而PCA聚类分析不能作为脓肿分枝杆菌脓肿亚种和马赛亚种的鉴别手段。Objective To perform the identification at the subspecies-level of Mycobacterium abscessus(M.abscessus)and analyze its characteristics based on matrix-assisted laser desorption ionization time-of-flight mass spectrometry(MALDI-TOF MS).Methods Biotyper software was used to construct the predicted peak spectrum of M.abscessus subsp.abscessus and M.abscessus subsp.massiliense.The predicted peak spectrum was constructed with expected maximum peak value number of 70 and peak frequency of 100%in the experimental group and control group,respectively.A blind test was performed on 31 strains of M.abscessus that were not used to construct predictive peak spectra to evaluate the identification efficiency of predictive peak spectra.FlexAnalysis software was used to summarize and analyze the list of mass spectral peak value of M.abscessus,and screen the specific peaks in mass spectra of different subspecies of M.abscessus.The principal component analysis(PCA)algorithm was used to perform the cluster analysis for the data from mass spectrometry of M.abscessus,and explore the feasibility of PCA clustering in distinguishing the subspecies of M.abscessus.Results In the experimental group,96.8%(30/31)of the strains were correctly identified,and one strain of M.abscessus subsp.massiliense with rough colony form was mistakenly identified as M.abscessus subsp.abscessus.In control group,77.4%(24/31)of the strains were correctly identified,but 7 strains of M.abscessus subsp.massiliense were incorrectly identified or unable to be identified.The identification efficiency in the experimental group was significantly better than that in the control group with statistical difference(χ^(2)=5.167,P=0.026).M.abscessus subsp.abscessus exhibited three specific peaks(m/z 4001.67,4386.81 and 4963.17),and M.abscessus subsp.massiliense also exhibited three specific peaks(m/z 4950.48,4381.78 and 5214.90).In the PCA 3D scatter plot,the data points of M.abscessus subsp.abscessus and M.abscessus subsp.massiliense were relatively dispersed without obviou

关 键 词：基质辅助激光解吸电离飞行时间质谱 脓肿分枝杆菌 菌种鉴定 主成分分析 

分 类 号：R372[医药卫生—病原生物学]