{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,26]],"date-time":"2026-08-26T10:27:47Z","timestamp":1787740067405,"version":"build-2784847793"},"reference-count":32,"publisher":"Wiley","issue":"8","license":[{"start":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T00:00:00Z","timestamp":1785369600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T00:00:00Z","timestamp":1785369600000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100002322","name":"Coordena\u00e7\u00e3o de Aperfei\u00e7oamento de Pessoal de N\u00edvel Superior","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100002322","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Current Protocols"],"published-print":{"date-parts":[[2026,8]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    The identification of mycobacteria remains challenging due to the high species diversity within the genus and the lack of a molecular method able to identify them in a single assay. Matrix\u2010assisted laser desorption\/ionization time\u2010of\u2010flight mass spectrometry (MALDI\u2010TOF MS) has been used as a rapid and promising alternative for bacterial identification; however, its reliability is highly dependent on effective protein extraction. This article describes a protocol specifically developed for the identification and species\u2010level discrimination of\u00a0\n                    <jats:italic>Mycobacterium<\/jats:italic>\n                    \u00a0spp. that integrates thermal inactivation, multiple biomass washes, mechanical disruption with zirconia beads, protein extraction using formic acid and acetonitrile, quality control, and troubleshooting. This procedure is compatible with Bruker MALDI Biotyper systems and is designed to meet biosafety level (BSL)\u20102 and BSL\u20103 laboratory requirements, ensuring operator safety and preventing aerosol generation. Emphasis is placed on standardizing critical steps, reproducibility, and safe handling of pathogenic strains. This protocol can be applied in both diagnostic and research laboratories, offering a reliable and standardized approach to preparing mycobacterial samples for analysis by MALDI\u2010TOF MS. \u00a9 2026 The Author(s).\n                    <jats:italic>Current Protocols<\/jats:italic>\n                    published by Wiley Periodicals LLC.\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol<\/jats:bold>\n                    : Protein extraction from\n                    <jats:italic>Mycobacterium<\/jats:italic>\n                    spp. cultured on solid media for identification by MALDI\u2010TOF mass spectrometry\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Support Protocol<\/jats:bold>\n                    : Preparation and quality control of mycobacterial cultures for MALDI\u2010TOF MS analysis\n                  <\/jats:p>","DOI":"10.1002\/cpz1.70425","type":"journal-article","created":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T14:51:42Z","timestamp":1785423102000},"source":"Crossref","is-referenced-by-count":0,"title":["Protein Extraction From\n                    <i>Mycobacterium<\/i>\n                    spp. for Identification by MALDI\u2010TOF Mass Spectrometry"],"prefix":"10.1002","volume":"6","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2597-5149","authenticated-orcid":false,"given":"Lucas Evangelista","family":"Marques","sequence":"first","affiliation":[{"name":"Department of Microbiology, Immunology and Parasitology Federal University of S\u00e3o Paulo  S\u00e3o Paulo Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stephanie Chrystine Balestro","family":"Mota","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Immunology and Parasitology Federal University of S\u00e3o Paulo  S\u00e3o Paulo Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christoffel","family":"Opperman","sequence":"additional","affiliation":[{"name":"South African Medical Research Council Centre for Tuberculosis Research, Division of Molecular Biology and Human Genetics, Faculty of Medicine and Health Sciences Stellenbosch University  Cape Town South Africa"},{"name":"National Health Laboratory Service Green Point TB\u2010Laboratory  Cape Town South Africa"},{"name":"Division of Medical Microbiology, Department of Pathology University of Cape Town  Cape Town South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Robin Mark","family":"Warren","sequence":"additional","affiliation":[{"name":"South African Medical Research Council Centre for Tuberculosis Research, Division of Molecular Biology and Human Genetics, Faculty of Medicine and Health Sciences Stellenbosch University  Cape Town South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Emilyn Costa","family":"Concei\u00e7\u00e3o","sequence":"additional","affiliation":[{"name":"South African Medical Research Council Centre for Tuberculosis Research, Division of Molecular Biology and Human Genetics, Faculty of Medicine and Health Sciences Stellenbosch University  Cape Town South Africa"},{"name":"Centre for Epidemic Response and Innovation (CERI), School for Data Science and Computational Thinking Stellenbosch University  Cape Town South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1068-7884","authenticated-orcid":false,"given":"Cristina","family":"Viana\u2010Niero","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Immunology and Parasitology Federal University of S\u00e3o Paulo  S\u00e3o Paulo Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,7,30]]},"reference":[{"key":"e_1_2_8_2_1_1","doi-asserted-by":"publisher","DOI":"10.5578\/mb.20229802"},{"key":"e_1_2_8_2_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cmi.2017.11.012"},{"key":"e_1_2_8_2_3_1","doi-asserted-by":"publisher","DOI":"10.3390\/microorganisms11071692"},{"key":"e_1_2_8_2_4_1","first-page":"1","volume-title":"Mycobacteria Extraction (MycoEX) Method: Standard Operating Procedure","author":"Bruker Daltonics","year":"2014"},{"key":"e_1_2_8_2_5_1","unstructured":"Bruker Daltonics. 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