Improved MALDI-TOF MS identification of Mycobacterium tuberculosis by use of an enhanced cell disruption protocol.
ABSTRACT - Mycobacterium tuberculosis is the microorganism that causes tuberculosis, a disease affecting millions of people worldwide. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) is a fast, reliable, and cost-effective method for microorganism identification which has been used for the identification of Mycobacterium spp. isolates. However, the mycobacteria cell wall is rich in lipids, which makes it difficult to obtain proteins for MALDI-TOF MS analysis. In this study, two cell preparation protocols were compared: the MycoEx, recommended by MALDI-TOF instrument manufacturer Bruker Daltonics, and the MycoLyser protocol described herein, which used the MagNA Lyser instrument to enhance cell disruption with ethanol. Cell disruption and protein extraction steps with the two protocols were performed using the Mycobacterium tuberculosis H37Rv strain, and the MALDI-TOF MS results were compared. The MycoLyser protocol allowed for improved Biotyper identification of M. tuberculosis since the log(score) values obtained with this protocol were mostly > 1.800 and significantly higher than that underwent MycoEx processing. The identification reliability was increased as well, considering the Bruker criteria. In view of these results, it is concluded that the MycoLyser protocol for mycobacterial cell disruption and protein extraction improves the MALDI-TOF MS method's efficacy for M. tuberculosis identification.
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Format: | Artigo de periódico biblioteca |
Language: | Ingles English |
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2023-09-05
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Subjects: | MALDI-TOF MS, Espectrometria, Microrganismo, Tuberculose, Mass spectrometry, Mycobacterium tuberculosis, Microorganisms, |
Online Access: | http://www.alice.cnptia.embrapa.br/alice/handle/doc/1156432 https://doi.org/10.3390/ microorganisms11071692 |
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dig-alice-doc-11564322023-09-05T17:23:40Z Improved MALDI-TOF MS identification of Mycobacterium tuberculosis by use of an enhanced cell disruption protocol. BACANELLI, G. ARAUJO, F. R. VERBISCK, N. V. GISELE BACANELLI, UNIVERSIDADE FEDERAL DE MATO GROSSO DO SUL; FLABIO RIBEIRO DE ARAUJO, CNPGC; NEWTON VALERIO VERBISCK, CNPGC. MALDI-TOF MS Espectrometria Microrganismo Tuberculose Mass spectrometry Mycobacterium tuberculosis Microorganisms ABSTRACT - Mycobacterium tuberculosis is the microorganism that causes tuberculosis, a disease affecting millions of people worldwide. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) is a fast, reliable, and cost-effective method for microorganism identification which has been used for the identification of Mycobacterium spp. isolates. However, the mycobacteria cell wall is rich in lipids, which makes it difficult to obtain proteins for MALDI-TOF MS analysis. In this study, two cell preparation protocols were compared: the MycoEx, recommended by MALDI-TOF instrument manufacturer Bruker Daltonics, and the MycoLyser protocol described herein, which used the MagNA Lyser instrument to enhance cell disruption with ethanol. Cell disruption and protein extraction steps with the two protocols were performed using the Mycobacterium tuberculosis H37Rv strain, and the MALDI-TOF MS results were compared. The MycoLyser protocol allowed for improved Biotyper identification of M. tuberculosis since the log(score) values obtained with this protocol were mostly > 1.800 and significantly higher than that underwent MycoEx processing. The identification reliability was increased as well, considering the Bruker criteria. In view of these results, it is concluded that the MycoLyser protocol for mycobacterial cell disruption and protein extraction improves the MALDI-TOF MS method's efficacy for M. tuberculosis identification. Communication. 2023-09-05T17:23:40Z 2023-09-05T17:23:40Z 2023-09-05 2023 Artigo de periódico Microorganisms, v. 11, issue 7, article 1692, 2023. http://www.alice.cnptia.embrapa.br/alice/handle/doc/1156432 https://doi.org/10.3390/ microorganisms11071692 Ingles en openAccess |
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MALDI-TOF MS Espectrometria Microrganismo Tuberculose Mass spectrometry Mycobacterium tuberculosis Microorganisms MALDI-TOF MS Espectrometria Microrganismo Tuberculose Mass spectrometry Mycobacterium tuberculosis Microorganisms |
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MALDI-TOF MS Espectrometria Microrganismo Tuberculose Mass spectrometry Mycobacterium tuberculosis Microorganisms MALDI-TOF MS Espectrometria Microrganismo Tuberculose Mass spectrometry Mycobacterium tuberculosis Microorganisms BACANELLI, G. ARAUJO, F. R. VERBISCK, N. V. Improved MALDI-TOF MS identification of Mycobacterium tuberculosis by use of an enhanced cell disruption protocol. |
description |
ABSTRACT - Mycobacterium tuberculosis is the microorganism that causes tuberculosis, a disease affecting millions of people worldwide. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) is a fast, reliable, and cost-effective method for microorganism identification which has been used for the identification of Mycobacterium spp. isolates. However, the mycobacteria cell wall is rich in lipids, which makes it difficult to obtain proteins for MALDI-TOF MS analysis. In this study, two cell preparation protocols were compared: the MycoEx, recommended by MALDI-TOF instrument manufacturer Bruker Daltonics, and the MycoLyser protocol described herein, which used the MagNA Lyser instrument to enhance cell disruption with ethanol. Cell disruption and protein extraction steps with the two protocols were performed using the Mycobacterium tuberculosis H37Rv strain, and the MALDI-TOF MS results were compared. The MycoLyser protocol allowed for improved Biotyper identification of M. tuberculosis since the log(score) values obtained with this protocol were mostly > 1.800 and significantly higher than that underwent MycoEx processing. The identification reliability was increased as well, considering the Bruker criteria. In view of these results, it is concluded that the MycoLyser protocol for mycobacterial cell disruption and protein extraction improves the MALDI-TOF MS method's efficacy for M. tuberculosis identification. |
author2 |
GISELE BACANELLI, UNIVERSIDADE FEDERAL DE MATO GROSSO DO SUL; FLABIO RIBEIRO DE ARAUJO, CNPGC; NEWTON VALERIO VERBISCK, CNPGC. |
author_facet |
GISELE BACANELLI, UNIVERSIDADE FEDERAL DE MATO GROSSO DO SUL; FLABIO RIBEIRO DE ARAUJO, CNPGC; NEWTON VALERIO VERBISCK, CNPGC. BACANELLI, G. ARAUJO, F. R. VERBISCK, N. V. |
format |
Artigo de periódico |
topic_facet |
MALDI-TOF MS Espectrometria Microrganismo Tuberculose Mass spectrometry Mycobacterium tuberculosis Microorganisms |
author |
BACANELLI, G. ARAUJO, F. R. VERBISCK, N. V. |
author_sort |
BACANELLI, G. |
title |
Improved MALDI-TOF MS identification of Mycobacterium tuberculosis by use of an enhanced cell disruption protocol. |
title_short |
Improved MALDI-TOF MS identification of Mycobacterium tuberculosis by use of an enhanced cell disruption protocol. |
title_full |
Improved MALDI-TOF MS identification of Mycobacterium tuberculosis by use of an enhanced cell disruption protocol. |
title_fullStr |
Improved MALDI-TOF MS identification of Mycobacterium tuberculosis by use of an enhanced cell disruption protocol. |
title_full_unstemmed |
Improved MALDI-TOF MS identification of Mycobacterium tuberculosis by use of an enhanced cell disruption protocol. |
title_sort |
improved maldi-tof ms identification of mycobacterium tuberculosis by use of an enhanced cell disruption protocol. |
publishDate |
2023-09-05 |
url |
http://www.alice.cnptia.embrapa.br/alice/handle/doc/1156432 https://doi.org/10.3390/ microorganisms11071692 |
work_keys_str_mv |
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