毛霉目病原真菌的实验室鉴定及体外抗真菌药物敏感性分析  

Laboratory identification and in vitro antifungal susceptibility analysis of the Mucorales

作　　者：袁凯旋[1] 叶静如 禄梦笛 陈卓曦 凌勇[1] 黄爱伟[1] 刘素玲[1] 顾兵 YUAN Kaixuan;YE Jingru;LU Mengdi;CHEN Zhuoxi;LING Yong;HUANG Aiwei;LIU Suling;GU Bing(Guangdong Provincial People′s Hospital/Guangdong Academy of Medical Sciences,Guangzhou 510080,Guangdong;School of Medical Technology,Guangdong Medical University,Dongguan 523808,Guangdong,China)

机构地区：[1]广东省人民医院/广东省医学科学院,广州510080 [2]广东医科大学医学技术学院,广东东莞523808

出　　处：《临床检验杂志》2022年第11期835-840,共6页Chinese Journal of Clinical Laboratory Science

摘　　要：目的以分子测序为参考,评估形态学、基质辅助激光解吸电离飞行时间质谱对毛霉目真菌的鉴定能力,分析毛霉目真菌对两性霉素B、泊沙康唑的体外药敏特点。方法收集2018年1月—2022年2月广东省人民医院25例毛霉病住院患者标本,进行镜检并对培养的菌落行形态学鉴定、质谱鉴定、分子测序和体外药敏试验。结果KOH湿片法阳性率(76%)高于革兰染色法(32%),差异有统计学意义(P<0.05);形态学鉴定可将68%的菌株鉴定到属水平,3株鉴定错误,2株无法鉴定;将所有菌株进行质谱鉴定,单用IVD和RUO数据库鉴定率分别为56%和44%,两库联合可将鉴定率提高为64%。毛霉目真菌对两性霉素B的抑制50%菌株生长的最小抑菌浓度(MIC_(50))为1μg/mL,根霉属相比其他属对两性霉素B表现出较高的MIC值,其中2株MIC>32μg/mL,另2株MIC为8μg/mL;根霉属对泊沙康唑的MIC_(50)为0.5μg/mL,横梗霉属MIC_(50)为4μg/mL,小克银汉霉属MIC_(50)为2μg/mL,有部分菌株表现出较低MIC值,同样亦有部分菌株对泊沙康唑表现出较高的MIC值。结论将传统镜检、培养与质谱技术和分子技术相结合,尽量将毛霉目真菌准确鉴定到种,并积极开展毛霉体外药敏试验,可为临床治疗提供参考依据。Objective To evaluate the identification ability of morphology and matrix-assisted laser desorption ionization-time of flight mass spectrometry(MALDI-TOF MS)for the Mucorales by using molecular sequencing as a reference,and analyze the susceptibility of Mucorales to amphotericin B and posaconazole in vitro.Methods Twenty-five samples of patients with mucormycosis hospitalized in Guangdong Provincial People′s Hospital between January 2018 and February 2022 were collected for microscopic examination,and the cultured colonies were performed morphological identification,mass spectrometry identification,molecular sequencing and in vitro antifungal susceptibility test.Results The positive rate of the KOH method(76%)was significantly higher than that of the Gram staining method(32%,P<0.05).Morphological identification could identify 68%of strains to the genus level.Three strains were identified incorrectly,and 2 strains could not be identified.All strains were identified by mass spectrometry,and the identification rates of IVD and RUO databases were 56%and 44%,respectively.The identification rate could be increased to 64%by combining the two databases.The MIC_(50)of the Mucorales to amphotericin B was 1μg/mL.The Rhizopus showed higher MIC value to amphotericin B than other genera.The MIC of 2 strains was>32μg/mL,and that of the other two strains was 8μg/mL.The MIC_(50)of the Rhizopus to posaconazole was 0.5μg/mL,and those of the Lichtheimia and Cunninghamia were 4μg/mL and 2μg/mL,respectively.Some strains showed lower MIC values to posaconazole,and some strains showed higher MIC values.Conclusion The combination of traditional microscopy,culture,mass spectrometry and molecular technology,accurate identification of species of the Mucorales as far as possible,and the active development of in vitro drug sensitivity tests of the Mucorales can provide reference for clinical treatment.

关 键 词：毛霉目 分子测序 形态学鉴定 基质辅助激光解吸电离飞行时间质谱 抗真菌体外药敏试验 

分 类 号：R446.5[医药卫生—诊断学]