{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,31]],"date-time":"2026-10-31T15:01:48Z","timestamp":1793458908448,"version":"4.5.0"},"reference-count":27,"publisher":"Springer Science and Business Media LLC","issue":"30","license":[{"start":{"date-parts":[[2025,11,5]],"date-time":"2025-11-05T00:00:00Z","timestamp":1762300800000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2025,11,5]],"date-time":"2025-11-05T00:00:00Z","timestamp":1762300800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"name":"German Federal Ministry of Economics and Climate Action","award":["13N13698"],"award-info":[{"award-number":["13N13698"]}]},{"DOI":"10.13039\/501100005713","name":"Technische Universit\u00e4t M\u00fcnchen","doi-asserted-by":"crossref","id":[{"id":"10.13039\/501100005713","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Anal Bioanal Chem"],"published-print":{"date-parts":[[2025,12]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    Chemiluminescence imaging of a\u00a0sandwich microarray immunoassay (CL-SMIA) is a simple and rapid method for antibody-based serotyping of pathogens like\n                    <jats:italic>L. pneumophila<\/jats:italic>\n                    , which is important to identify even outbreak-associated subtypes like\n                    <jats:italic>L. pneumophila<\/jats:italic>\n                    Sg 1 subtype Philadelphia. The CL image pattern can be interpreted by a decision path similar to the well-known \u201cDresden Panel\u201d, which is typically performed via indirect ELISA. New antibodies needed to be characterized for the so-called LegioTyper panel as capturing antibodies to perform the flow-based CL-SMIA. The following was achieved for the LegioTyper principle: (i) minimizing the costs for antibody microarrays by manufacturing Jeffamine-2003-coated polycarbonate sheets, (ii) optimized automated test procedure which is able to perform the CL-SMIA in 34\u00a0min, and (iii) covalent immobilization of cAb using a microdispensing buffer which contains reactions for stabilizing and bioconjugation of cAb. Nine mAb and one polyclonal Ab (pAb) were characterized regarding the selectivity to\n                    <jats:italic>L. pneumophila<\/jats:italic>\n                    Sg 1 and additionally to the virulence-associated subtypes Knoxville, France, Allentown, Benidorm, and Philadelphia (all Pontiac group), OLDA, Oxford, Bellingham, Heysham, and Camperdown (all non-Pontiac group). We could show that by using the LegioTyper it is possible to differentiate between\n                    <jats:italic>L. pneumophila<\/jats:italic>\n                    Sg1 and non-Sg1 and between Pontiac and non-Pontiac strains. Additionally, subtyping with the \u201cLegioTyper Panel\u201d was possible for most of the outbreak-associated subtypes. Overall, our study showed that the LegioTyper principle can be used for rapid antibody-based subtyping of\n                    <jats:italic>L. pneumophila<\/jats:italic>\n                    Sg 1 after cultivation. It is an alternative to indirect ELISA or qPCR, which are more labor- and cost-intensive, and the automation is more complicated.\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Graphical Abstract<\/jats:bold>\n                  <\/jats:p>","DOI":"10.1007\/s00216-025-06194-3","type":"journal-article","created":{"date-parts":[[2025,11,5]],"date-time":"2025-11-05T09:12:05Z","timestamp":1762333925000},"page":"6797-6809","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["LegioTyper: Rapid typing of Legionella pneumophila serogroup 1 by flow-based chemiluminescence sandwich microarray immunoassay"],"prefix":"10.1007","volume":"417","author":[{"given":"C.","family":"B\u00e4rwinkel","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"A.","family":"Petzold","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"C.","family":"L\u00fcck","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"M.","family":"Petzold","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"M.","family":"Seidel","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,11,5]]},"reference":[{"key":"6194_CR1","doi-asserted-by":"publisher","DOI":"10.1093\/femsre\/fuaf022","volume":"49","author":"F Hammes","year":"2025","unstructured":"Hammes F, Gabrielli M, Cavallaro A, Eichelberg A, Barigelli S, Bigler M, et al. Foresight 2035: a perspective on the next decade of research on the management of Legionella spp. in engineered aquatic environments. FEMS Microbiol Rev. 2025;49: fuaf022. https:\/\/doi.org\/10.1093\/femsre\/fuaf022.","journal-title":"FEMS Microbiol Rev"},{"key":"6194_CR2","doi-asserted-by":"publisher","first-page":"882","DOI":"10.1111\/jam.12559","volume":"117","author":"E Heijnsbergen","year":"2014","unstructured":"Heijnsbergen E, Roda Husman AM, Lodder WJ, Bouwknegt M, Docters Van Leeuwen AE, Bruin JP, et al. Viable Legionella pneumophila bacteria in natural soil and rainwater puddles. J Appl Microbiol. 2014;117:882\u201390. https:\/\/doi.org\/10.1111\/jam.12559.","journal-title":"J Appl Microbiol"},{"key":"6194_CR3","doi-asserted-by":"publisher","first-page":"21660","DOI":"10.1016\/j.bioflm.2024.100196","volume":"14","author":"A Barbosa","year":"2024","unstructured":"Barbosa A, Azevedo NF, Goeres DM, Cerqueira L. Ecology of Legionella pneumophila biofilms: the link between transcriptional activity and the biphasic cycle. Biofilm. 2024;14:21660\u201375. https:\/\/doi.org\/10.1016\/j.bioflm.2024.100196.","journal-title":"Biofilm"},{"key":"6194_CR4","doi-asserted-by":"publisher","DOI":"10.1080\/21505594.2021.1903199","volume":"12","author":"D Chauhan","year":"2021","unstructured":"Chauhan D, Shames SR. Pathogenicity and virulence of Legionella: intracellular replication and host response. Virulence. 2021;12: 1. https:\/\/doi.org\/10.1080\/21505594.2021.1903199.","journal-title":"Virulence"},{"issue":"4","key":"6194_CR5","doi-asserted-by":"publisher","DOI":"10.1128\/iai.00441-22","volume":"91","author":"SR Shames","year":"2023","unstructured":"Shames SR. Eat or be eaten: strategies used by Legionella to acquire host-derived nutrients and evade lysosomal degradation. Infect Immun. 2023;91(4): e00441-22. https:\/\/doi.org\/10.1128\/iai.00441-22.","journal-title":"Infect Immun"},{"issue":"3","key":"6194_CR6","doi-asserted-by":"publisher","DOI":"10.3390\/pathogens13030250","volume":"13","author":"S Cristino","year":"2024","unstructured":"Cristino S, Pascale MR, Marino F, Derelitto C, Salaris S, Orsini M, Squarzoni S, Grottola A, Girolamini L. Characterization of a novel species of Legionella isolated from a healthcare facility: Legionella resiliens sp. nov. Pathogens. 2024;13(3): 250. https:\/\/doi.org\/10.3390\/pathogens13030250.","journal-title":"Pathogens"},{"key":"6194_CR7","doi-asserted-by":"publisher","first-page":"51","DOI":"10.1186\/s13613-024-01252-y","volume":"14","author":"J Rello","year":"2024","unstructured":"Rello J, Allam C, Ruiz-Spinelli A, Jarraud S. Severe legionnaires\u2019 disease. 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All patients had provided written informed consent for the use of their clinical data for research purposes as part of the institutional Broad Consent framework at the University Hospital Dresden.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics approval"}},{"value":"All samples were collected with informed consent.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Consent to participate"}},{"value":"The authors declare no competing interests.","order":4,"name":"Ethics","group":{"name":"EthicsHeading","label":"Conflict of interest"}}]}}