Identification of filamentous fungi based on morphological features is the most available approach used in clinical mycology laboratories. However, MALDI-TOF mass spectrometry is currently invaluable for identification of microorganisms because of its rapidity, simplicity, and accuracy. This study aimed to find the optimal way of identifying filamentous fungi using MALDI-TOF MS.

The sample comprised 193 isolates of filamentous fungi. The identification started with morphological assessment. Then isolates were identified using MALDI-TOF MS, both directly from culture and following culture in liquid media with extraction. Subsequently, identification of 20 selected isolates was compared by sequencing of the benA gene, ITS1-5,8-ITS2, and D1-D2 LSU regions.

Based on morphological criteria, 17 genera of fungi were identified. With MALDI-TOF MS performed directly from culture, nine isolates were identified to the genus level and 184 to the species level, with a total of 75 species being noted. With the MALDI-TOF MS extraction method, 190 isolates were identified to the species level, with 43 species being noted. The rates of agreement between identification using morphology and the MALDI-TOF MS direct method were 58.55% at the genus level and 22.24% at the species level. The rates of agreement between identification using morphology and the MALDI-TOF MS extraction method were 84.97% at the genus level and 46.11% at the species level. Using sequencing, 87.5% agreement was found for identification with the MALDI-TOF MS extraction method, as compared with only 43.75% for the direct method.

The results suggest that the optimal approach to identification of filamentous fungi is a combination of morphological features and MALDI-TOF MS using the extraction method.

基于形态特征鉴定丝状真菌是临床真菌学实验室最常用的方法。然而，MALDI-TOF 质谱法由于其快速、简单和准确，目前在微生物鉴定方面具有不可估量的价值。本研究旨在寻找使用 MALDI-TOF MS 鉴定丝状真菌的最佳方法。

样本包括 193 个分离的丝状真菌。鉴定从形态学评估开始。然后使用 MALDI-TOF MS 鉴定分离物，直接来自培养物和随后在液体培养基中培养并提取。随后，通过对benA基因、ITS1-5,8-ITS2 和 D1-D2 LSU 区域进行测序，比较了 20 个选定分离株的鉴定。

根据形态学标准，鉴定出17个真菌属。使用直接从培养物中进行的 MALDI-TOF MS，9 个分离株被鉴定到属水平，184 个分离到种水平，总共有 75 个物种被记录。使用 MALDI-TOF MS 提取方法，190 个分离株被鉴定到物种水平，其中 43 个物种被记录。形态学鉴定与MALDI-TOF MS直接法鉴定在属水平上的一致性为58.55%，在种水平上为22.24%。形态学鉴定与MALDI-TOF MS提取法在属水平上的一致性为84.97%，在种水平上为46.11%。使用测序，发现与 MALDI-TOF MS 提取方法鉴定的一致性为 87.5%，而直接方法仅为 43.75%。

结果表明，鉴定丝状真菌的最佳方法是结合形态特征和MALDI-TOF MS提取法。