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Identification of Nocardia species using matrix-assisted laser desorption/ionization–time-of-flight mass spectrometry
BACKGROUND: The MALDI (matrix-assisted laser desorption/ionization) Biotyper system for bacterial identification has already been utilized in clinical microbiology laboratories as a successful clinical application of protoemics. However, in cases of Nocardia, mass spectra suitable for MALDI Biotyper...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4409724/ https://www.ncbi.nlm.nih.gov/pubmed/25931991 http://dx.doi.org/10.1186/s12014-015-9078-5 |
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author | Segawa, Shunsuke Nishimura, Motoi Sogawa, Kazuyuki Tsuchida, Sachio Murata, Shota Watanabe, Masaharu Matsushita, Kazuyuki Kamei, Katsuhiko Nomura, Fumio |
author_facet | Segawa, Shunsuke Nishimura, Motoi Sogawa, Kazuyuki Tsuchida, Sachio Murata, Shota Watanabe, Masaharu Matsushita, Kazuyuki Kamei, Katsuhiko Nomura, Fumio |
author_sort | Segawa, Shunsuke |
collection | PubMed |
description | BACKGROUND: The MALDI (matrix-assisted laser desorption/ionization) Biotyper system for bacterial identification has already been utilized in clinical microbiology laboratories as a successful clinical application of protoemics. However, in cases of Nocardia, mass spectra suitable for MALDI Biotyper identification are often not obtained if such specimens are processed like general bacteria. This problem is related to the insufficiencies in bacterial spectrum databases that preclude accurate specimen identification. Here, we developed a bacterial processing method to improve mass spectra from specimens of the genus Nocardia. In addition, with the new processing method, we constructed a novel in-house bacterial database that combines a commercial database and mass spectra of Nocardia strains from the Department of Clinical Laboratory at Chiba University Hospital (DCLC) and the Medical Mycology Research Center at Chiba University (MMRC). RESULTS: The newly developed method (Nocardia Extraction Method at DCLC [NECLC]) based on ethanol-formic acid extraction (EFAE) improved mass spectra obtained from Nocardia specimens. The Nocardia in-house database at Chiba University Hospital (NDCUH) was then successfully validated. In brief, prior to introduction of the NECLC and NDCUH, 10 of 64 (15.6%) clinical isolates were identified at the species level and 16 isolates (25.0%) could only be identified at the genus level. In contrast, after the introduction, 58 isolates (90.6%) were identified at the species level and 6 isolates (9.4%) were identified at the genus level. CONCLUSIONS: The results of this study suggest that MALDI-TOF (time-of-flight) Biotyper system can identify Nocardia accurately in a short time in combination with a simple processing method and an in-house database. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12014-015-9078-5) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-4409724 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-44097242015-04-30 Identification of Nocardia species using matrix-assisted laser desorption/ionization–time-of-flight mass spectrometry Segawa, Shunsuke Nishimura, Motoi Sogawa, Kazuyuki Tsuchida, Sachio Murata, Shota Watanabe, Masaharu Matsushita, Kazuyuki Kamei, Katsuhiko Nomura, Fumio Clin Proteomics Research BACKGROUND: The MALDI (matrix-assisted laser desorption/ionization) Biotyper system for bacterial identification has already been utilized in clinical microbiology laboratories as a successful clinical application of protoemics. However, in cases of Nocardia, mass spectra suitable for MALDI Biotyper identification are often not obtained if such specimens are processed like general bacteria. This problem is related to the insufficiencies in bacterial spectrum databases that preclude accurate specimen identification. Here, we developed a bacterial processing method to improve mass spectra from specimens of the genus Nocardia. In addition, with the new processing method, we constructed a novel in-house bacterial database that combines a commercial database and mass spectra of Nocardia strains from the Department of Clinical Laboratory at Chiba University Hospital (DCLC) and the Medical Mycology Research Center at Chiba University (MMRC). RESULTS: The newly developed method (Nocardia Extraction Method at DCLC [NECLC]) based on ethanol-formic acid extraction (EFAE) improved mass spectra obtained from Nocardia specimens. The Nocardia in-house database at Chiba University Hospital (NDCUH) was then successfully validated. In brief, prior to introduction of the NECLC and NDCUH, 10 of 64 (15.6%) clinical isolates were identified at the species level and 16 isolates (25.0%) could only be identified at the genus level. In contrast, after the introduction, 58 isolates (90.6%) were identified at the species level and 6 isolates (9.4%) were identified at the genus level. CONCLUSIONS: The results of this study suggest that MALDI-TOF (time-of-flight) Biotyper system can identify Nocardia accurately in a short time in combination with a simple processing method and an in-house database. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12014-015-9078-5) contains supplementary material, which is available to authorized users. BioMed Central 2015-03-07 /pmc/articles/PMC4409724/ /pubmed/25931991 http://dx.doi.org/10.1186/s12014-015-9078-5 Text en © Segawa et al.; licensee BioMed Central. 2015 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Research Segawa, Shunsuke Nishimura, Motoi Sogawa, Kazuyuki Tsuchida, Sachio Murata, Shota Watanabe, Masaharu Matsushita, Kazuyuki Kamei, Katsuhiko Nomura, Fumio Identification of Nocardia species using matrix-assisted laser desorption/ionization–time-of-flight mass spectrometry |
title | Identification of Nocardia species using matrix-assisted laser desorption/ionization–time-of-flight mass spectrometry |
title_full | Identification of Nocardia species using matrix-assisted laser desorption/ionization–time-of-flight mass spectrometry |
title_fullStr | Identification of Nocardia species using matrix-assisted laser desorption/ionization–time-of-flight mass spectrometry |
title_full_unstemmed | Identification of Nocardia species using matrix-assisted laser desorption/ionization–time-of-flight mass spectrometry |
title_short | Identification of Nocardia species using matrix-assisted laser desorption/ionization–time-of-flight mass spectrometry |
title_sort | identification of nocardia species using matrix-assisted laser desorption/ionization–time-of-flight mass spectrometry |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4409724/ https://www.ncbi.nlm.nih.gov/pubmed/25931991 http://dx.doi.org/10.1186/s12014-015-9078-5 |
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