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Current hospital imaging protocols often delay treatment, underscoring the need for portable diagnostic solutions. We have investigated silver nanostars (AgNS) incubated with human plasma, deposited on a simple aluminum foil substrate, and utilizing Surface-Enhanced Raman Spectroscopy (SERS) combined with machine learning (ML) to provide a proof-of-concept for rapid differentiation of stroke types. These are the seminal steps for the development of low-cost pre-hospital diagnostics at point-of-care, with potential for improving patient outcomes. The proposed SERS assay aims to classify plasma from stroke patients, differentiating hemorrhagic from ischemic stroke. Silver nanostars were incubated with plasma and spiked with glial fibrillary acidic protein (GFAP), a biomarker elevated in hemorrhagic stroke. SERS spectra were analyzed using ML to distinguish between hemorrhagic and ischemic stroke, mimicked by different concentrations of GFAP. Key innovations include optimized AgNS\u2013plasma incubates formation, controlled plasma-to-AgNS ratios, and a low-cost aluminum foil substrate, enabling results within 15 min. Differential analysis revealed stroke-specific protein profiles, while ML improved classification accuracy through ensemble modeling and feature engineering. The integrated ML model achieved rapid and precise stroke predictions within seconds, demonstrating the assay\u2019s potential for immediate clinical decision-making.<\/jats:p>","DOI":"10.3390\/bios15030136","type":"journal-article","created":{"date-parts":[[2025,2,24]],"date-time":"2025-02-24T05:36:47Z","timestamp":1740375407000},"page":"136","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Towards Rapid and Low-Cost Stroke Detection Using SERS and Machine Learning"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0009-0007-7128-1427","authenticated-orcid":false,"given":"Cristina","family":"Freitas","sequence":"first","affiliation":[{"name":"Associate Laboratory i4HB\u2014Institute for Health and Bioeconomy, Faculdade de Ci\u00eancias e Tecnologia, Universidade NOVA de Lisboa, 2819-516 Caparica, Portugal"},{"name":"UCIBIO\u2014Applied Molecular Biosciences Unit, Departamento de Qu\u00edmica, Faculdade de Ci\u00eancias e Tecnologia, Universidade NOVA de Lisboa, 2819-516 Caparica, Portugal"}]},{"given":"Jo\u00e3o","family":"Eleut\u00e9rio","sequence":"additional","affiliation":[{"name":"COPELABS\u2014Departamento de Engenharia Inform\u00e1tica e Sistemas de Informa\u00e7\u00e3o, Universidade Lus\u00f3fona, Centro Universit\u00e1rio de Lisboa, 1749-024 Lisboa, Portugal"}]},{"given":"Gabriela","family":"Soares","sequence":"additional","affiliation":[{"name":"COPELABS\u2014Departamento de Engenharia Inform\u00e1tica e Sistemas de Informa\u00e7\u00e3o, Universidade Lus\u00f3fona, Centro Universit\u00e1rio de Lisboa, 1749-024 Lisboa, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5429-5874","authenticated-orcid":false,"given":"Maria","family":"Enea","sequence":"additional","affiliation":[{"name":"LAQV\/REQUIMTE\u2014Laborat\u00f3rio Associado para a Qu\u00edmica Verde\/Rede de Qu\u00edmica e Tecnologia, Departamento de Qu\u00edmica e Bioqu\u00edmica, Faculdade de Ci\u00eancias, Universidade do Porto, 4169-007 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3115-6588","authenticated-orcid":false,"given":"Daniela","family":"Nunes","sequence":"additional","affiliation":[{"name":"Associate Laboratory i3N, Departamento de Ci\u00eancia dos Materiais, Faculdade de Ci\u00eancias e Tecnologia, Universidade NOVA de Lisboa, and CEMOP\/UNINOVA, 2829-516 Caparica, Portugal"}]},{"given":"Elvira","family":"Fortunato","sequence":"additional","affiliation":[{"name":"Associate Laboratory i3N, Departamento de Ci\u00eancia dos Materiais, Faculdade de Ci\u00eancias e Tecnologia, Universidade NOVA de Lisboa, and CEMOP\/UNINOVA, 2829-516 Caparica, Portugal"}]},{"given":"Rodrigo","family":"Martins","sequence":"additional","affiliation":[{"name":"Associate Laboratory i3N, Departamento de Ci\u00eancia dos Materiais, Faculdade de Ci\u00eancias e Tecnologia, Universidade NOVA de Lisboa, and CEMOP\/UNINOVA, 2829-516 Caparica, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7350-649X","authenticated-orcid":false,"given":"Hugo","family":"\u00c1guas","sequence":"additional","affiliation":[{"name":"Associate Laboratory i3N, Departamento de Ci\u00eancia dos Materiais, Faculdade de Ci\u00eancias e Tecnologia, Universidade NOVA de Lisboa, and CEMOP\/UNINOVA, 2829-516 Caparica, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2086-5696","authenticated-orcid":false,"given":"Eul\u00e1lia","family":"Pereira","sequence":"additional","affiliation":[{"name":"LAQV\/REQUIMTE\u2014Laborat\u00f3rio Associado para a Qu\u00edmica Verde\/Rede de Qu\u00edmica e Tecnologia, Departamento de Qu\u00edmica e Bioqu\u00edmica, Faculdade de Ci\u00eancias, Universidade do Porto, 4169-007 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9415-3742","authenticated-orcid":false,"given":"Helena L. A.","family":"Vieira","sequence":"additional","affiliation":[{"name":"Associate Laboratory i4HB\u2014Institute for Health and Bioeconomy, Faculdade de Ci\u00eancias e Tecnologia, Universidade NOVA de Lisboa, 2819-516 Caparica, Portugal"},{"name":"UCIBIO\u2014Applied Molecular Biosciences Unit, Departamento de Qu\u00edmica, Faculdade de Ci\u00eancias e Tecnologia, Universidade NOVA de Lisboa, 2819-516 Caparica, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0235-2531","authenticated-orcid":false,"given":"L\u00facio Studer","family":"Ferreira","sequence":"additional","affiliation":[{"name":"COPELABS\u2014Departamento de Engenharia Inform\u00e1tica e Sistemas de Informa\u00e7\u00e3o, Universidade Lus\u00f3fona, Centro Universit\u00e1rio de Lisboa, 1749-024 Lisboa, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5139-2871","authenticated-orcid":false,"given":"Ricardo","family":"Franco","sequence":"additional","affiliation":[{"name":"Associate Laboratory i4HB\u2014Institute for Health and Bioeconomy, Faculdade de Ci\u00eancias e Tecnologia, Universidade NOVA de Lisboa, 2819-516 Caparica, Portugal"},{"name":"UCIBIO\u2014Applied Molecular Biosciences Unit, Departamento de Qu\u00edmica, Faculdade de Ci\u00eancias e Tecnologia, Universidade NOVA de Lisboa, 2819-516 Caparica, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2025,2,22]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"e102","DOI":"10.1212\/WNL.0b013e3182a94f3c","article-title":"Stroke, tPA, and Physician Decision-Making","volume":"81","author":"Hovsepian","year":"2013","journal-title":"Neurology"},{"key":"ref_2","unstructured":"(2024, December 30). 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