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MALDI-TOF MS直接涂靶法快速鉴定丝状真菌的临床应用
引用本文:张桂,王丽赟,王玫,顾海彤,鲁辛辛. MALDI-TOF MS直接涂靶法快速鉴定丝状真菌的临床应用[J]. 疾病监测, 2019, 34(11): 980-986. DOI: 10.3784/j.issn.1003-9961.2019.11.008
作者姓名:张桂  王丽赟  王玫  顾海彤  鲁辛辛
作者单位:首都医科大学附属北京同仁医院,北京,100730
基金项目:首都医科大学附属北京同仁医院科研骨干培育基金(No. 2014-YJJ-GGL-038);国家科技重大专项(No. 2014ZX10004005)
摘    要:目的评价基质辅助激光解吸电离飞行时间质谱直接涂靶法(质谱直涂法)快速鉴定丝状真菌的临床应用价值。方法对2010年1月至2017年12月首都医科大学附属北京同仁医院临床标本中分离的133株丝状真菌分别进行形态学、基因序列分析及质谱直涂法鉴定,并将鉴定结果进行分析比较。采用SPSS 16.0软件进行统计分析,计数资料采用χ^2检验。结果以基因序列分析结合形态学鉴定的结果为标准,将133株丝状真菌鉴定到种、复合群和属的鉴定率:质谱直涂法分别为64.66%、79.70%和96.24%,基因序列分析则分别为69.17%、83.46%和99.25%,而形态学鉴定分别为68.42%、84.21%和93.23%。质谱直涂法与另外两种方法的种、复合群和属的鉴定率的差异无统计学意义(χ^2值分别为0.611、0.625、2.728和0.516、0.915、1.206,P值均>0.05)。质谱直涂法将77株曲霉属中61.04%、88.31%和98.70%的菌株分别鉴定到种、复合群和属,将14株青霉属中64.29%和92.86%的菌株分别鉴定到种和属,还将42株其他丝状真菌中的71.43%和92.86%的菌株分别鉴定到种和属;因数据库中真菌种类不足,致3.76%(5/133)的真菌无效鉴定。结论MALDI-TOF MS直接涂靶法鉴定丝状真菌的能力可与形态学鉴定和基因序列分析相媲美,因其快速且操作简便,临床应用值得推广。

关 键 词:直接涂靶  丝状真菌  快速鉴定  基质辅助激光解吸电离飞行时间质谱技术
收稿时间:2019-03-11

Evaluation of the clinical application of MALDI-TOF MS with directly smeared target in rapid identification of filamentous fungi
Affiliation:Beijing Tongren Hospital, Capital Medical University, Beijing 100730, China
Abstract:ObjectiveTo evaluate the clinical application of matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) with directly smeared target (MS direct smear method) in the rapid identification of filamentous fungi.MethodsA study was performed on 133 strains of filamentous fungi cultured from clinical samples in Beijing Tongren Hospital affiliated to Capital Medical University from January 2010 to December 2017, and these strains were identified by morphological observation, gene sequence analysis and MS direct smear method, respectively. The results were analyzed and compared. χ2 test was performed in the enumeration data analysis by using software SPSS 16.0.ResultsBased on the results of 133 strains of filamentous fungi identified by gene sequence analysis and morphological observation, the identification rates of species, complex and genera of the filamentous fungi strains by MS direct smear method were 64.66%, 79.70% and 96.24%, respectively, while the rates obtained by gene sequence analysis were 69.17%, 83.46% and 99.25%, respectively, and then the rates by morphological identification were 68.42%, 84.21% and 93.23%, respectively. Furthermore, there were no significant differences in the identification rates of the species, complexes and genera between MS direct smear method and latter two methods (χ2=0.611, 0.625, 2.728, respectively, and χ2=0.516, 0.915, 1.206, respectively, P>0.05). MS direct smear method identified 61.04%, 88.31% and 98.70% of the 77 strains of Aspergillus, respectively, at species, complex and genera levels, 64.29% and 92.86% of the 14 strains of Penicillium at species and genera levels, and 71.43% and 92.86% of 42 other filamentous fungi strains at species and genera levels. Five strains (3.76%, 5/133) were not identified due to the insufficient species of fungi in the database.ConclusionMALDI-TOF MS with directly smeared target can accurately identify clinical filamentous fungi with similar results to morphological observation and gene sequence analysis. It is the test which is rapid, easy and simple to handle for use in clinical practice.
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