基于基质辅助激光解吸电离飞行时间质谱的克柔念珠菌遗传分化研究  

Genetic differentiation of Candida krusei based on matrix-assisted laser desorption ionization time-of-flight mass spectrometry

作　　者：龚杰 张沈茜 张慧芳[1] 肖盟[3,4] 何利华 赵飞[1] 徐英春[3,4] 张建中[1] 肖迪 GongJie;Zhang Shenxi;Zhang Huifang;Xiao Meng;He Lihua;Zhao Fei;Xu Yingchun;Zhang Jianzhong;Xiao Di(State Key Laboratory fo r Communicable Disease Prevention and Control,National Institute for Communicable Disease Control and Prevention,Chinese Center for Disease Control and Prevention,Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases,Beijing 102206,China;Department of Dermatology,Peking University International Hospital,Beijing 102206,China;Department of Clinical Laboratory,Peking Union Medical College Hospital,Chinese Academy of Medical Sciences,Beijing 100730,China;Beijing Key Laboratory for Mechanisms Research and Precision Diagnosis of Invasive Fungal Diseases,Beijing 100730,China)

机构地区：[1]中国疾病预防控制中心传染病预防控制所,传染病预防控制国家重点实验室,感染性疾病诊治协同创新中心,北京102206 [2]北京大学国际医院皮肤科,北京102206 [3]中国医学科学院北京协和医院检验科,北京100730 [4]“侵袭性真菌病机制研究与精准诊断”北京市重点实验室,北京100730

出　　处：《疾病监测》2019年第11期969-973,共5页Disease Surveillance

基　　金：国家科技重大专项（No.2018ZX10712001,No.2018ZX10733402）;中国疾病预防控制中心青年科研基金（No.2018A102）~~

摘　　要：目的在蛋白质水平明确克柔念珠菌的遗传分化特征,建立基于基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)的不同谱系克柔念珠菌的鉴定方法。方法选取20株克柔念珠菌,采用MALDI-TOF MS结合分析软件ClinProTools,确定克柔念珠菌不同谱系的质谱标志峰,构建基于遗传算法(GA)的分类模型,用于区分不同谱系的克柔念珠菌菌株,选择25株克柔念珠菌对该模型进行验证。结果利用20株克柔念珠菌构建的GA分类模型,反映模型正确鉴别能力的识别能力值(RC)为100%,反映模型处理测试谱图变异能力的交叉验证值(CV)为97.89%。25株克柔念珠菌、150张谱图外部验证模型显示其分类能力为95.30%。确定13个克柔念珠菌谱系相关的特征峰(m/z 3971.40、3136.95、3427.33、2405.28、2996.73、2913.95、3376.97、6736.13、5819.03、4045.16、5869.00、3618.10及3946.14),可用于正确区分来自不同谱系的克柔念珠菌。结论本研究论证并确认了克柔念珠菌的群体遗传分化事件,同时也建立了一种简便而有效的快速谱系分型方法。Objective To identify the genetic differentiation characteristics of Candida krusei at the level of protein and establish different lineage identification methods based on matrix-assisted laser desorption ionization time-of-flight mass spectrometry(MALDI-TOF MS).Methods MALDI-TOF MS coupled to ClinProTools was used to discover MALDI-TOF MS biomarker peaks and establish a classification model based on a genetic algorithm(GA)to differentiate the different lineages of C.krusei.Twenty strains of C.krusei were used to establish an analysis model,and 25 strains of were used for the validation.Results For the GA typing model using 20 strains of C.krusei,the recognition capability(RC)value,which reflects the model’s ability to correctly identify its component spectra was 100%,and the cross-validation(CV)value,which reflects the ability of the model to handle variability among the test spectra was 97.89%and the classification power value of the validation model for 25 C.krusei strains and 150 mass spectrums was 95.30%.This model contained 13 biomarker peaks(m/z 3971.40,3136.95,3427.33,2405.28,2996.73,2913.95,3376.97,6736.13,5819.03,4045.16,5869.00,3618.10 and3946.14)and can be used to correctly identify different lineages of C.krusei.Conclusion This study demonstrated not only the population genetic differentiation of C.krusei from the peptide level,but also confirmed the existence of this event,and more importantly,a simple and effective rapid lineage typing method was established in this study.

关 键 词：克柔念珠菌 谱系 遗传分化 基质辅助激光解吸电离飞行时间质谱 分类模型 

分 类 号：R379.4[医药卫生—病原生物学]