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

Ya-Ting Ning; Ya-Ting Ning; Ya-Ting Ning; Wen-Hang Yang; Wen-Hang Yang; Wen-Hang Yang; Wei Zhang; Wei Zhang; Meng Xiao; Meng Xiao; Meng Xiao; Yao Wang; Yao Wang; Jing-Jia Zhang; Jing-Jia Zhang; Ge Zhang; Ge Zhang; Si-Meng Duan; Si-Meng Duan; Ai-Ying Dong; Da-Wen Guo; Gui-Ling Zou; Hai-Nan Wen; Yan-Yan Guo; Li-Ping Chen; Miao Chai; Jing-Dong He; Qiong Duan; Li-Xia Zhang; Li Zhang; Li Zhang; Ying-Chun Xu; Ying-Chun Xu

doi:10.3389/fcimb.2021.687240

Frontiers in Cellular and Infection Microbiology (Jul 2021)

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

Ya-Ting Ning,
Ya-Ting Ning,
Ya-Ting Ning,
Wen-Hang Yang,
Wen-Hang Yang,
Wen-Hang Yang,
Wei Zhang,
Wei Zhang,
Meng Xiao,
Meng Xiao,
Meng Xiao,
Yao Wang,
Yao Wang,
Jing-Jia Zhang,
Jing-Jia Zhang,
Ge Zhang,
Ge Zhang,
Si-Meng Duan,
Si-Meng Duan,
Ai-Ying Dong,
Da-Wen Guo,
Gui-Ling Zou,
Hai-Nan Wen,
Yan-Yan Guo,
Li-Ping Chen,
Miao Chai,
Jing-Dong He,
Qiong Duan,
Li-Xia Zhang,
Li Zhang,
Li Zhang,
Ying-Chun Xu,
Ying-Chun Xu

Affiliations

Ya-Ting Ning: Department of Clinical Laboratory, State Key Laboratory of Complex Severe and Rare Diseases, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China
Ya-Ting Ning: Graduate School, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China
Ya-Ting Ning: Beijing Key Laboratory for Mechanisms Research and Precision Diagnosis of Invasive Fungal Diseases, Beijing, China
Wen-Hang Yang: Department of Clinical Laboratory, State Key Laboratory of Complex Severe and Rare Diseases, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China
Wen-Hang Yang: Graduate School, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China
Wen-Hang Yang: Beijing Key Laboratory for Mechanisms Research and Precision Diagnosis of Invasive Fungal Diseases, Beijing, China
Wei Zhang: Graduate School, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China
Wei Zhang: Clinical Microbiology Laboratory, The First Affiliated Hospital of Hebei North University, Zhangjiakou, China
Meng Xiao: Department of Clinical Laboratory, State Key Laboratory of Complex Severe and Rare Diseases, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China
Meng Xiao: Graduate School, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China
Meng Xiao: Beijing Key Laboratory for Mechanisms Research and Precision Diagnosis of Invasive Fungal Diseases, Beijing, China
Yao Wang: Department of Clinical Laboratory, State Key Laboratory of Complex Severe and Rare Diseases, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China
Yao Wang: Beijing Key Laboratory for Mechanisms Research and Precision Diagnosis of Invasive Fungal Diseases, Beijing, China
Jing-Jia Zhang: Department of Clinical Laboratory, State Key Laboratory of Complex Severe and Rare Diseases, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China
Jing-Jia Zhang: Beijing Key Laboratory for Mechanisms Research and Precision Diagnosis of Invasive Fungal Diseases, Beijing, China
Ge Zhang: Department of Clinical Laboratory, State Key Laboratory of Complex Severe and Rare Diseases, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China
Ge Zhang: Beijing Key Laboratory for Mechanisms Research and Precision Diagnosis of Invasive Fungal Diseases, Beijing, China
Si-Meng Duan: Department of Clinical Laboratory, State Key Laboratory of Complex Severe and Rare Diseases, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China
Si-Meng Duan: Beijing Key Laboratory for Mechanisms Research and Precision Diagnosis of Invasive Fungal Diseases, Beijing, China
Ai-Ying Dong: Department of Clinical Laboratory, North China University of Science and Technology Affiliated Hospital, Tangshan, China
Da-Wen Guo: Department of Clinical Laboratory, The First Affiliated Hospital of Harbin Medical University, Harbin, China
Gui-Ling Zou: Department of Clinical Laboratory, The Fourth Affiliated Hospital of Harbin Medical University, Harbin, China
Hai-Nan Wen: Department of Laboratory, The Affiliated Hospital of Chengde Medical University, Chengde, China
Yan-Yan Guo: Department of Clinical Laboratory, Tangshan Worker’s Hospital, Tangshan, China
Li-Ping Chen: 0Department of Laboratory Medicine, Mudanjiang First People’s Hospital, Heilongjiang, China
Miao Chai: 1Department of Clinical Laboratory, The First Hospital of Harbin, Harbin, China
Jing-Dong He: 2Department of Clinical Laboratory, Tianjin Chest Hospital, Tianjin, China
Qiong Duan: 3Department of Clinical Laboratory, Jinling Province People’s Hospital, Jinling, China
Li-Xia Zhang: 4Department of Clinical Laboratory, Shanxi Provincial People’s Hospital, Taiyuan, China
Li Zhang: Department of Clinical Laboratory, State Key Laboratory of Complex Severe and Rare Diseases, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China
Li Zhang: Beijing Key Laboratory for Mechanisms Research and Precision Diagnosis of Invasive Fungal Diseases, Beijing, China
Ying-Chun Xu: Department of Clinical Laboratory, State Key Laboratory of Complex Severe and Rare Diseases, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China
Ying-Chun Xu: Beijing Key Laboratory for Mechanisms Research and Precision Diagnosis of Invasive Fungal Diseases, Beijing, China

DOI: https://doi.org/10.3389/fcimb.2021.687240
Journal volume & issue: Vol. 11

Abstract

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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 overall mean of maximum and minimum S/N ratio higher than that by FUS. An in-house library of M-Discover MS was successfully built from 135 isolates from 42 species belonging to 18 genera using the ZSB method. Analysis of 467 isolates resulted in 97.22% correctly identified isolates to the species level by the ZSB method versus 95.50% by the routine method. The two novel methods are time- and cost-effective and allow efficient identification of filamentous fungi while providing a simplified procedure to build an in-house library. Thus, more clinical laboratories may consider adopting MALDI-TOF MS for filamentous fungi identification in the future.

Published in Frontiers in Cellular and Infection Microbiology

ISSN: 2235-2988 (Online)
Publisher: Frontiers Media S.A.
Country of publisher: Switzerland
LCC subjects: Science: Microbiology
Website: http://www.frontiersin.org/Cellular-and-Infection-Microbiology

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