Department of Medical Microbiology, Postgraduate Institute of Medical Education and Research, Chandigarh, India.
Jhaveri Microbiology Centre, Brien Holden Eye Research Centre, L V Prasad Eye Institute, Hyderabad, India.
Proteomics Clin Appl. 2019 May;13(3):e1800070. doi: 10.1002/prca.201800070. Epub 2018 Sep 24.
Invasive fungal infections caused by melanized fungi are a growing concern. Rapid and reliable identification plays an important role in optimizing therapy. Matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF MS)-based identification has emerged as a faster and more accurate diagnostic technique. However, lack of a protein extraction protocol and limited database restricts the identification of melanized fungi by MALDI-TOF MS. The study is designed to standardize protein extraction protocol, to enrich the existing, and to create an in-house database for the rapid identification of melanized fungi.
In this study, 59 sequence-confirmed, melanized fungi were used to expand and to create an in-house database using a modified protein extraction protocol. A total of 117 clinical isolates are further used to validate the created database.
Using existing Bruker database, only 29(24.8%) out of 117 moulds could be identified. However, all the isolates are identified accurately by supplementing the Bruker database with the created in-house database. MALDI-TOF MS takes significantly lesser time for identification compared to DNA sequencing.
An expanded database with modified protein extraction protocol can reduce significant time to identify melanized fungi by MALDI-TOF MS.
由黑真菌引起的侵袭性真菌感染是一个日益受到关注的问题。快速而可靠的鉴定在优化治疗中起着重要作用。基于基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF MS)的鉴定已成为一种更快、更准确的诊断技术。然而,缺乏蛋白质提取方案和有限的数据库限制了 MALDI-TOF MS 对黑真菌的鉴定。本研究旨在标准化蛋白质提取方案,丰富现有的数据库,并创建一个内部数据库,以快速鉴定黑真菌。
在这项研究中,使用了 59 株经序列确认的黑真菌,使用改良的蛋白质提取方案进行扩展和创建内部数据库。进一步使用总共 117 株临床分离株来验证创建的数据库。
使用现有的 Bruker 数据库,仅能鉴定出 117 株霉菌中的 29 株(24.8%)。然而,通过用创建的内部数据库补充 Bruker 数据库,可以准确地鉴定出所有分离株。MALDI-TOF MS 进行鉴定的时间明显短于 DNA 测序。
通过 MALDI-TOF MS 鉴定黑真菌时,使用改良的蛋白质提取方案的扩展数据库可以显著缩短鉴定所需的时间。