{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,8]],"date-time":"2025-10-08T15:43:28Z","timestamp":1759938208993,"version":"3.41.2"},"reference-count":56,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2025,2,4]],"date-time":"2025-02-04T00:00:00Z","timestamp":1738627200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Cell. Infect. Microbiol."],"abstract":"<jats:p>Two factors frequently impede accurate bacterial identification using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS): inadequate bacterial abundance in real samples and bacterial combinations. For MALDI-TOF MS analysis and libraries for bacterial identification, time-consuming culture procedures are necessary to achieve sufficient concentration and isolation of a single bacterium. When dealing with hazardous bacteria like <jats:italic>Brucella<\/jats:italic>, which are more difficult to handle and cure, this problem becomes even more crucial. To overcome these obstacles, Fe<jats:sub>3<\/jats:sub>O<jats:sub>4<\/jats:sub> magnetic nanoparticles (MNPs) linked with <jats:italic>Brucella<\/jats:italic>-specific antibodies and MALDI-TOF MS analysis have been used to create a quick and accurate technique for direct bacterial separation and identification in complex samples. This method allows MNPs to immune-selectively collect <jats:italic>Brucella<\/jats:italic> cells, which are then deactivated and ready for MALDI-TOF MS analysis by a formic acid\/acetonitrile wash. Rabbits were used to manufacture brucella antibodies, which have effectively adsorbed onto the MNPs\u2013protein A. Any particular <jats:italic>Brucella<\/jats:italic> bacteria found in the media might be absorbed by this MNPs\u2013protein A\u2013antibody immunoprobe. The concentration of <jats:italic>Brucella<\/jats:italic> bacterial cells increases the protein spectrum\u2019s visibility by a factor of 10<jats:sup>3<\/jats:sup>, making it possible to quickly identify <jats:italic>Brucella<\/jats:italic> spp. without first growing them in cultural conditions. This method has been successfully used to achieve a limit of detection (LOD) of 50 CFU\/mL in an aqueous medium and genuine sample\u2014milk. The diagnostic time for this harmful bacterium is greatly decreased because the entire procedure from bacterial isolation to species identification is finished in less than 60 min. High sensitivity and specificity are demonstrated by the immunoassay\u2013MS approach, as the spectral pattern it produces matches well-known databases like SPECLUST and Ribopeaks.<\/jats:p>","DOI":"10.3389\/fcimb.2025.1531018","type":"journal-article","created":{"date-parts":[[2025,2,4]],"date-time":"2025-02-04T06:33:37Z","timestamp":1738650817000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":1,"title":["Immunoassay\u2013mass spectrometry to identify Brucella melitensis"],"prefix":"10.3389","volume":"15","author":[{"given":"Amirreza","family":"Sharif","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ramin Bagheri","family":"Nejad","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alireza","family":"Ghassempour","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1965","published-online":{"date-parts":[[2025,2,4]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"3693","DOI":"10.1038\/s41598-024-54236-0","article-title":"Automated sepsis detection with vancomycin-and allantoin-polydopamine magnetic nanoparticles","volume":"14","author":"Abafogi","year":"2024","journal-title":"Sci. Rep."},{"key":"B2","doi-asserted-by":"publisher","first-page":"2739","DOI":"10.1002\/pmic.201200053","article-title":"Nonculture-based identification of bacteria in milk by protein fingerprinting","volume":"12","author":"Barreiro","year":"2012","journal-title":"Proteomics"},{"key":"B3","doi-asserted-by":"publisher","first-page":"143","DOI":"10.1038\/s41579-021-00621-9","article-title":"Mass spectrometry-based metabolomics in microbiome investigations","volume":"20","author":"Bauermeister","year":"2022","journal-title":"Nat. Rev. Microbiol."},{"key":"B4","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2023.1204452","article-title":"MALDI-TOF MS in microbiological diagnostics: future applications beyond identification","volume":"14","author":"Becker","year":"2023","journal-title":"Front. Microbiol."},{"key":"B5","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1186\/s12889-020-09016-6","article-title":"Milk-borne bacterial health hazards in milk produced for commercial purpose in Tigray, northern Ethiopia","volume":"20","author":"Berhe","year":"2020","journal-title":"BMC Public Health"},{"key":"B6","doi-asserted-by":"publisher","first-page":"114234","DOI":"10.1016\/j.bios.2022.114234","article-title":"Microneedle patches integrated with lateral flow cassettes for blood-free chronic kidney disease point-of-care testing during a pandemic","volume":"208","author":"Chen","year":"2022","journal-title":"Biosens. Bioelectron."},{"key":"B7","doi-asserted-by":"publisher","first-page":"346","DOI":"10.1002\/prca.201500086","article-title":"Recent development of mass spectrometry and proteomics applications in identification and typing of bacteria","volume":"10","author":"Cheng","year":"2016","journal-title":"PROTEOMICS\u2013Clinical Appl."},{"key":"B8","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2024.1443636","article-title":"Insights in infectious agents and disease: 2022","volume":"15","author":"Cloeckaert","year":"2024","journal-title":"Front. Microbiol."},{"key":"B9","doi-asserted-by":"publisher","DOI":"10.3389\/fvets.2022.1009928","article-title":"MALDI-TOF mass spectrometry profiling of bovine skim milk for subclinical mastitis detection","volume":"9","author":"Cuccato","year":"2022","journal-title":"Front. Vet. Sci."},{"key":"B10","doi-asserted-by":"publisher","DOI":"10.3389\/fcimb.2024.1379188","article-title":"Foodborne bacterial pathogens under the One Health perspective-antimicrobial resistance, epidemiology, virulence, and zoonotic impact","volume":"14","author":"de Oliveira","year":"2024","journal-title":"Front. Cell. Infection Microbiol."},{"key":"B11","doi-asserted-by":"publisher","first-page":"989","DOI":"10.15212\/ZOONOSES-2021-0028","article-title":"The animal origin of major human infectious diseases: what can past epidemics teach us about preventing the next pandemic","volume":"2","author":"Dharmarajan","year":"2022","journal-title":"Zoonoses"},{"key":"B12","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2023.1193841","article-title":"Community series-innovative approaches in diagnosis of emerging\/re-emerging infectious diseases, volume II","volume":"14","author":"Foster","year":"2023","journal-title":"Front. Microbiol."},{"key":"B13","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2016.01891","article-title":"Non-selective separation of bacterial cells with magnetic nanoparticles facilitated by varying surface charge","volume":"7","author":"Gao","year":"2016","journal-title":"Front. Microbiol."},{"key":"B14","doi-asserted-by":"publisher","DOI":"10.3389\/fphar.2024.1397602","article-title":"Improving the treatment of bacterial infections caused by multidrug-resistant bacteria through drug repositioning","volume":"15","author":"Glajzner","year":"2024","journal-title":"Front. Pharmacol."},{"key":"B15","doi-asserted-by":"publisher","first-page":"351","DOI":"10.1007\/s13206-022-00064-1","article-title":"Recent developments in innovative magnetic nanoparticles-based immunoassays: from improvement of conventional immunoassays to diagnosis of COVID-19","volume":"16","author":"Ha","year":"2022","journal-title":"BioChip J."},{"key":"B16","doi-asserted-by":"publisher","first-page":"80","DOI":"10.3390\/microbiolres14010008","article-title":"The current level of MALDI-TOF MS applications in the detection of microorganisms: a short review of benefits and limitations","volume":"14","author":"Haider","year":"2023","journal-title":"Microbiol. Res. (Pavia)."},{"key":"B17","doi-asserted-by":"publisher","first-page":"1530","DOI":"10.1021\/jasms.2c00110","article-title":"A combination of MALDI-TOF MS proteomics and species-unique biomarkers\u2019 discovery for rapid screening of brucellosis","volume":"33","author":"Hamidi","year":"2022","journal-title":"J. Am. Soc Mass Spectrom."},{"key":"B18","doi-asserted-by":"publisher","first-page":"297","DOI":"10.1007\/s13534-021-00202-y","article-title":"Recent advances in magnetic nanoparticle-based microfluidic devices for the pretreatment of pathogenic bacteria","volume":"11","author":"Han","year":"2021","journal-title":"Biomed. Eng. Lett."},{"key":"B19","doi-asserted-by":"publisher","DOI":"10.1021\/acsami.4c11169","article-title":"Bacterial binding to polydopamine-coated magnetic nanoparticles","author":"Houser","year":"2024","journal-title":"ACS Appl. Mater. Interfaces"},{"key":"B20","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2024.1428304","article-title":"Rapid identification of bloodstream infection pathogens and drug resistance using Raman spectroscopy enhanced by convolutional neural networks","volume":"15","author":"Kang","year":"2024","journal-title":"Front. Microbiol."},{"key":"B21","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2023.1273364","article-title":"Microbe-nanoparticle interactions: a mechanistic approach","volume":"14","author":"Kumari","year":"2023","journal-title":"Front. Microbiol."},{"key":"B22","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2018.01229","article-title":"Cultivable microbial diversity associated with cellular phones","volume":"9","author":"Kurli","year":"2018","journal-title":"Front. Microbiol."},{"key":"B23","doi-asserted-by":"publisher","DOI":"10.3389\/fcimb.2022.1079184","article-title":"Performance evaluation and clinical validation of optimized nucleotide MALDI-TOF-MS for mycobacterial identification","volume":"12","author":"Li","year":"2022","journal-title":"Front. Cell. Infect. Microbiol."},{"key":"B24","doi-asserted-by":"publisher","DOI":"10.3389\/fvets.2022.827702","article-title":"MALDI-TOF mass spectrometry as a rapid screening alternative for non-tuberculous mycobacterial species identification in the veterinary laboratory","volume":"9","author":"Lorente-Leal","year":"2022","journal-title":"Front. Vet. Sci."},{"key":"B25","doi-asserted-by":"publisher","first-page":"173","DOI":"10.1002\/ejic.200400504","article-title":"Rearrangement of the active ester intermediate during HOBt\/EDC amide coupling","volume":"2005","author":"Mahmoud","year":"2005","journal-title":"Eur. J. Inorg. Chem."},{"key":"B26","doi-asserted-by":"publisher","first-page":"2300093","DOI":"10.1002\/masy.202300093","article-title":"Assessing the microbial contamination levels in milk samples from rural and urban areas: A focus on raisen and bhopal districts","volume":"413","author":"Mukhopadhyay","year":"2024","journal-title":"Macromol. Symp."},{"key":"B27","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2014.00005","article-title":"AAV vectors vaccines against infectious diseases","volume":"5","author":"Nieto","year":"2014","journal-title":"Front. Immunol."},{"key":"B28","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1186\/s12014-020-09278-7","article-title":"Mass spectrometry-based microbiological testing for blood stream infection","volume":"17","author":"Nomura","year":"2020","journal-title":"Clin. Proteomics"},{"key":"B29","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2013.00103","article-title":"The intrinsic resistome of bacterial pathogens","volume":"4","author":"Olivares","year":"2013","journal-title":"Front. Microbiol."},{"key":"B30","doi-asserted-by":"crossref","first-page":"2251","DOI":"10.3390\/diagnostics11122251","article-title":"Multicenter evaluation of rapid BACpro\u00ae II for the accurate identification of microorganisms directly from blood cultures using MALDI-TOF MS","volume":"11","author":"Ovia\u00f1o","year":"2021","journal-title":"Diagnostics"},{"key":"B31","doi-asserted-by":"publisher","first-page":"46","DOI":"10.2174\/1874318801004010046","article-title":"Diagnosis of brucellosis","volume":"4","author":"Padilla","year":"2010","journal-title":"Open Vet. Sci. J."},{"key":"B32","doi-asserted-by":"publisher","first-page":"120","DOI":"10.3390\/microorganisms11010120","article-title":"Evaluation of two different Preparation protocols for MALDI-TOF MS nontuberculous mycobacteria identification from Liquid and Solid Media","volume":"11","author":"Pastrone","year":"2023","journal-title":"Microorganisms"},{"key":"B33","doi-asserted-by":"publisher","DOI":"10.3389\/fcimb.2021.813133","article-title":"Progress in pathogen identification based on mass spectrometry","volume":"11","author":"Peng","year":"2022","journal-title":"Front. Cell. Infection Microbiol."},{"key":"B34","doi-asserted-by":"crossref","first-page":"1744","DOI":"10.3390\/life12111744","article-title":"Sepsityper\u00ae kit versus in-house method in rapid identification of bacteria from positive blood cultures by MALDI-TOF mass spectrometry","volume":"12","author":"Per\u0161e","year":"2022","journal-title":"Life"},{"key":"B35","doi-asserted-by":"publisher","first-page":"4444","DOI":"10.3168\/jds.2009-2182","article-title":"The effect of centrifugation and resuspension on the recovery of Mycoplasma species from milk","volume":"92","author":"Punyapornwithaya","year":"2009","journal-title":"J. Dairy Sci."},{"key":"B36","doi-asserted-by":"publisher","DOI":"10.3389\/fpubh.2023.1186800","article-title":"Case report: A case of brucellosis misdiagnosed as coronavirus disease 2019\/influenza in China","volume":"11","author":"Qin","year":"2023","journal-title":"Front. Public Heal."},{"key":"B37","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2019.02954","article-title":"MALDI-TOF MS application in microbial ecology studies","volume":"10","author":"Rahi","year":"2020","journal-title":"Front. Microbiol."},{"key":"B38","doi-asserted-by":"crossref","first-page":"123091","DOI":"10.1016\/j.jchromb.2021.123091","article-title":"Study of Glutathione S-transferase-P1 in cancer blood plasma after extraction by affinity magnetic nanoparticles and monitoring by MALDI-TOF, IM-Q-TOF and LC-ESI-Q-TOF MS","volume":"1190","author":"Ramandi","year":"2022","journal-title":"J. Chromatogr. B"},{"key":"B39","doi-asserted-by":"publisher","first-page":"321","DOI":"10.1002\/mas.21534","article-title":"Characterization of microbial mixtures by mass spectrometry","volume":"37","author":"Sandrin","year":"2018","journal-title":"Mass Spectrom. Rev."},{"key":"B40","doi-asserted-by":"publisher","first-page":"9623","DOI":"10.1021\/acs.analchem.9b01002","article-title":"Rapid bacteria detection at low concentrations using sequential immunomagnetic separation and paper-based isotachophoresis","volume":"91","author":"Schaumburg","year":"2019","journal-title":"Anal. Chem."},{"key":"B41","doi-asserted-by":"publisher","first-page":"13631","DOI":"10.3390\/ijms232113631","article-title":"Biomolecular profiling by MALDI-TOF mass spectrometry in food and beverage analyses","volume":"23","author":"\u0160ebela","year":"2022","journal-title":"Int. J. Mol. Sci."},{"key":"B42","doi-asserted-by":"publisher","first-page":"1140","DOI":"10.3390\/pr8091140","article-title":"Recent applications of magnetic nanoparticles in food analysis","volume":"8","author":"Socas-Rodr\u00edguez","year":"2020","journal-title":"Processes"},{"key":"B43","doi-asserted-by":"publisher","first-page":"10460","DOI":"10.1021\/acs.analchem.0c01119","article-title":"Conjugation of single-chain variable fragment antibody to magnetic nanoparticles and screening of fig mosaic virus by MALDI TOF mass spectrometry","volume":"92","author":"Soleimani Mashhadi","year":"2020","journal-title":"Anal. Chem."},{"key":"B44","doi-asserted-by":"publisher","first-page":"418","DOI":"10.3390\/nano10030418","article-title":"Proinflammatory effect of carbon-based nanomaterials: In vitro study on stimulation of inflammasome NLRP3 via destabilisation of lysosomes","volume":"10","author":"Svadlakova","year":"2020","journal-title":"Nanomaterials"},{"key":"B45","doi-asserted-by":"publisher","DOI":"10.1016\/j.heliyon.2024.e32020","article-title":"Nanotechnology improves the detection of bacteria: Recent advances and future perspectives","author":"Takallu","year":"2024","journal-title":"Heliyon"},{"key":"B46","doi-asserted-by":"publisher","first-page":"e00800","DOI":"10.1128\/mbio.00800-22","article-title":"Microbes and climate change: a research prospectus for the future","volume":"13","author":"Tiedje","year":"2022","journal-title":"MBio"},{"key":"B47","doi-asserted-by":"publisher","first-page":"2814","DOI":"10.3390\/app12062814","article-title":"MALDI-based mass spectrometry in clinical testing: Focus on bacterial identification","volume":"12","author":"Tsuchida","year":"2022","journal-title":"Appl. Sci."},{"key":"B48","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2016.00415","article-title":"Evaluation of the Bruker Biotyper matrix-assisted laser desorption\/ionization time-of-flight mass spectrometry system for identification of clinical and environmental isolates of Burkholderia pseudomallei","volume":"7","author":"Wang","year":"2016","journal-title":"Front. Microbiol."},{"key":"B49","doi-asserted-by":"publisher","DOI":"10.3389\/fbioe.2022.962746","article-title":"Detection nanodevices for infectious diseases","volume":"10","author":"Wang","year":"2022","journal-title":"Front. Bioeng. Biotechnol."},{"key":"B50","doi-asserted-by":"publisher","first-page":"9416","DOI":"10.1021\/acsami.6b16831","article-title":"Efficient enrichment and analyses of bacteria at ultralow concentration with quick-response magnetic nanospheres","volume":"9","author":"Wen","year":"2017","journal-title":"ACS Appl. Mater. Interfaces"},{"key":"B51","doi-asserted-by":"publisher","first-page":"107388","DOI":"10.1016\/j.microc.2022.107388","article-title":"Rapid enrichment and detection of Staphylococcus aureus in milk using polyethyleneimine functionalized magnetic nanoparticles","volume":"178","author":"Xiao","year":"2022","journal-title":"Microchem. J."},{"key":"B52","doi-asserted-by":"publisher","DOI":"10.3389\/fbioe.2019.00141","article-title":"Applications of iron oxide-based magnetic nanoparticles in the diagnosis and treatment of bacterial infections","volume":"7","author":"Xu","year":"2019","journal-title":"Front. Bioeng. Biotechnol."},{"key":"B53","doi-asserted-by":"publisher","first-page":"10","DOI":"10.1128\/CMR.00073-19","article-title":"Laboratory diagnosis of human brucellosis","volume":"33","author":"Yagupsky","year":"2019","journal-title":"Clin. Microbiol. Rev."},{"key":"B54","doi-asserted-by":"publisher","first-page":"10400","DOI":"10.1021\/acs.analchem.8b02258","article-title":"Direct MALDI-TOF MS identification of bacterial mixtures","volume":"90","author":"Yang","year":"2018","journal-title":"Anal. Chem."},{"key":"B55","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2024.1449844","article-title":"Artificial intelligence applications in the diagnosis and treatment of bacterial infections","volume":"15","author":"Zhang","year":"2024","journal-title":"Front. Microbiol."},{"key":"B56","doi-asserted-by":"publisher","DOI":"10.3389\/fvets.2023.1162465","article-title":"Recent trends in the use of bacteriophages as replacement of antimicrobials against food-animal pathogens","volume":"10","author":"Zia","year":"2023","journal-title":"Front. Vet. Sci."}],"container-title":["Frontiers in Cellular and Infection Microbiology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fcimb.2025.1531018\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,2,4]],"date-time":"2025-02-04T06:33:42Z","timestamp":1738650822000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fcimb.2025.1531018\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,2,4]]},"references-count":56,"alternative-id":["10.3389\/fcimb.2025.1531018"],"URL":"https:\/\/doi.org\/10.3389\/fcimb.2025.1531018","relation":{},"ISSN":["2235-2988"],"issn-type":[{"type":"electronic","value":"2235-2988"}],"subject":[],"published":{"date-parts":[[2025,2,4]]},"article-number":"1531018"}}