99株马尔尼菲篮状菌质谱聚类分析及体外药敏研究  

MALDI-TOF MS-based clustering and antifungal susceptibility of 99 Talaromyces marneffei strains

作　　者：黄春兰[1] 贺月园 梁辉苍 覃仁利 郭建[2] HUANG Chunlan;HE Yueyuan;LIANG Huicang;QIN Renli;GUO Jian(Liuzhou People's Hospital,Medical Laboratoy Department,Liuzhou 545001,Guangxi,China;Department of Laboratory Medicine,Shanghai East Hospital,Tongj University School of Medicine,Shanghai 200123)

机构地区：[1]柳州市人民医院检验科,广西柳州545001 [2]上海市东方医院南院医学检验科,上海200123

出　　处：《中国艾滋病性病》2023年第2期146-150,共5页Chinese Journal of Aids & STD

基　　金：柳州市科技局课题(2022CAC0215);广西壮族自治区卫生健康委员会重点实验室项目:广西真菌研究与真菌病防治重点实验室(ZZH2020004);广西医科大学第一附属医院重点实验室培育项目:广西亚热带真菌与真菌病研究重点实验室(YYZS2020006)。

摘　　要：目的 研究99株马尔尼菲篮状菌(TM)亲缘聚类分析关系及体外酵母相对10种常规抗真菌药物的最低抑菌浓度(MIC)。方法 采用基质辅助激光解析串联飞行时间质谱仪(MALDI-TOF MS)对TM进行鉴定、聚类分析,体外药敏试验采用37℃培养7~9天变成酵母相后,调整菌悬液为0.5麦氏浓度,接种到药敏板条上37℃孵育,检测MIC。结果 MALDI-TOF MS对TM鉴定平均得分(2.2±0.1)分,与形态学方法及rDNA-ITS测序结果相一致,聚类分析可将TM分为三大簇。药敏显示99株(100.0%)艾沙康唑的MIC均≤0.015μg/mL;伊曲康唑、伏立康唑和泊沙康唑MIC均≤0.03μg/mL;99株(100.0%)两性霉素B的MIC值≤0.5μg/mL,其中83株(83.8%)MIC≤0.25μg/mL;91株(91.9%)5-氟胞嘧啶MIC值≤0.5μg/mL;49株(49.5%)氟康唑的MIC值≤1μg/mL;56株(56.6%)卡泊芬净MIC值≤2μg/mL;70株(70.7%)米卡芬净MIC值≥8μg/mL;88株(88.9%)阿尼芬净MIC值≥2μg/mL。TM对艾沙康唑、伊曲康唑、伏立康唑、泊沙康唑、5-氟胞嘧啶和两性霉素B MIC处于低水平,棘白菌素类卡泊芬净、米卡芬净、阿尼芬净MIC值均处于高水平。结论 MALDI-TOF MS能够对TM进行准确鉴定和亲缘性聚类分析,根据TM体外的MIC提示敏感性最高依次是艾沙康唑、伊曲康唑、伏立康唑、泊沙康唑、两性霉素B,敏感性最低的是棘白菌素类三种抗真菌药。Objective To conduct cluster analysis and the minimum antibacterial concentration(MIC) of the 99Talaromyces marneffei strains of antifungal drugs in yeast. Methods MALDI-TOF MS was used to identified and cluster T.M isolates, all the strans incubated at 37 ℃ for 7 to 9 days to become yeast phase, adjusting the bacterial to 0.5(McF). Then we measured the minimum inhibitory concentration(MIC). Results The average score of MALDI-TOF MS was 2.2±0.1. The identification rate reached 100.0%. Hierarchical clustering of MALDI-TOF peak profiles identified three different clusters. The results suggested that 99(100.0%) strains against Isavuconazole with MIC≤0.015 μg/mL.Itraconazole, voriconazole, and posaconazole were all with MIC≤0.003 μg/mL. Amphotericin B with MIC was ≤0.5 μg/mL, of which 83 strains(83.8%) against amphotericin B with MIC≤0.25 μg/mL,91 strains(91.9%) against 5-flucytosine with MIC≤0.5 μg/mL,49(49.5%) strains against fluconazole with MIC≤1 μg/mL, 56 strains(56.6%) against caspofungin with MIC≤2 μg/mL, 70(70.7%) strains against micafungin with MIC≥8 μg/mL and 88 strains(88.9%) strains against anidulafungin with MIC≥2 μg/mL. The susceptibilities results indicated that isavuconazole, itraconazole, voriconazole, posaconazole, 5-fluorocytosine, and amphotericin B had higher activity against T. marneffei yeasts. Caspofungin, micafungin, and anidulafungin had the lowest activity. Conclusions MALDI-TOF MS can be considered as a method for identifiaution and clustering of Tmarneffei. The results of antifungal susceptibilities in vitro revealed that the highest activity was oxiconazole, itraconazole, voriconazole, posaconazole, amphotericin B, 5-fluorocytosine and echinocandins had the lower activity against T. marneffei yeasts.

关 键 词：马尔尼菲篮状菌 飞行时间质谱技术 聚类分析 最低抑菌浓度 

分 类 号：R373.9[医药卫生—病原生物学] R512.91[医药卫生—基础医学]