Developing Two Rapid Protein Extraction Methods Using Focused-Ultrasonication and Zirconia-Silica Beads for Filamentous Fungi Identification by MALDI-TOF MS

作者：Yating Ning, Wenhang Yang, Wei Zhang, Meng Xiao, Yao Wang, Jingjia Zhang, Ge Zhang, Simeng Duan, Aiying Dong, Dawen Guo, Gui-Ling Zou, Hainan Wen, Yan-Yan Guo, Liping Chen, Miao Chai, Jingdong He, Duan Qiong, Lixia Zhang, Lixia Zhang, Li Zhang, Li Zhang, Yingchun Xu · 发表于：Frontiers in Cellular and Infection Microbiology · 年份：2021 · DOI：10.3389/fcimb.2021.687240 · 被引用次数：26 · 研究领域：Bacterial Identification and Susceptibility Testing、Plant Pathogens and Fungal Diseases、Genomics and Phylogenetic Studies

Filamentous fungi identification by Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) has been challenging due to the lack of simple and rapid protein extraction methods and insufficient species coverage in the database. In this study, we created two rapid protein extraction methods for filamentous fungi: a one-step zirconia-silica beads method (ZSB) and a focused-ultrasonication method (FUS). The identification accuracy of two methods were evaluated with the VITEK MS, as well as number of spectra peaks and signal-to-noise ratio (S/N) with M-Discover 100 MALDI-TOF MS compared to the routine method. The better method was applied to build a filamentous fungi in-house spectra library for the M-Discover 100 MS, and then another one and routine method were performed in parallel to verify the accuracy and commonality of the in-house library. Using the two optimized methods, the dedicated operating time before MALDI-TOF MS analysis was reduced from 30 min to 7 (ZSB) or 5 (FUS) min per sample, with only a few seconds added for each additional strain. And both two methods identified isolates from most mold types equal to or better than the routine method, and the total correct identification rate using VITEK MS was 79.67, 76.42, and 76.42%, respectively. On the other hand, the two rapid methods generally achieved higher maximum and minimum S/N ratios with these isolates tested as compared to the routine method. Besides, the ZSB method produced...