Marinach-Patrice C, Lethuillier A, Marly A, Brossas J-Y, Gené J, Symoens F, Datry A, Guarro J, Mazier D, Hennequin C
Université Pierre et Marie Curie, INSERM, UMR S 945, APHP, Hôpital St Antoine, Service Parasitologie-Mycologie, Centre d'Investigations Biomedicales, Groupe hospitalier Pitié-Salpétrière, Paris, France.
Clin Microbiol Infect. 2009 Jul;15(7):634-42. doi: 10.1111/j.1469-0691.2009.02758.x. Epub 2009 May 18.
Fusarium spp. have recently emerged as significant human pathogens. Identification of these species is important, both for epidemiological purposes and for patient management, but conventional identification based on morphological traits is hindered by major phenotypic polymorphism. In this study, 62 strains, or isolates, belonging to nine Fusarium species were subjected to both molecular identification TEF1 gene sequencing and matrix-assisted laser desorption ionization-time-of-flight (MALDI-TOF) analysis. Following stringent standardization, the proteomic-based method appeared to be both reproducible and robust. Mass spectral analysis by comparison with a database, built in this study, of the most frequently isolated species, including Fusarium solani, Fusarium oxysporum, Fusarium verticilloides, Fusarium proliferatum and Fusarium dimerum, correctly identified 57 strains. As expected, the four species (i.e. Fusarium chlamydosporum, Fusarium equiseti, Fusarium polyphialidicum, Fusarium sacchari) not represented in the database were not identified. Results from mass spectrometry and molecular identification agreed in five of the six cases in which results from morphological and molecular identification were not in agreement. MALDI-TOF yielded results within 1 h, making it a valuable tool for identifying clinical Fusarium isolates at the species level. Uncommon species must now be added to the database. MALDI-TOF may also prove useful for identifying other clinically important moulds.
镰刀菌属最近已成为重要的人类病原体。对这些菌种进行鉴定,无论是出于流行病学目的还是患者管理目的都很重要,但基于形态特征的传统鉴定方法受到主要表型多态性的阻碍。在本研究中,对属于9种镰刀菌的62株菌株或分离株进行了分子鉴定(TEF1基因测序)和基质辅助激光解吸电离飞行时间(MALDI-TOF)分析。经过严格标准化后,基于蛋白质组学的方法似乎具有可重复性和稳健性。通过与本研究构建的最常分离菌种(包括茄病镰刀菌、尖孢镰刀菌、轮枝镰刀菌、层出镰刀菌和双隔镰刀菌)的数据库进行比较的质谱分析,正确鉴定了57株菌株。正如预期的那样,数据库中未包含的4种菌种(即厚垣镰刀菌、木贼镰刀菌、多瓶梗镰刀菌、甘蔗镰刀菌)未被鉴定出来。在形态学和分子鉴定结果不一致的6个案例中,有5个案例的质谱分析结果与分子鉴定结果一致。MALDI-TOF在1小时内得出结果,使其成为在菌种水平上鉴定临床镰刀菌分离株的有价值工具。现在必须将罕见菌种添加到数据库中。MALDI-TOF可能也被证明对鉴定其他临床上重要的霉菌有用。