{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T15:54:45Z","timestamp":1790610885778,"version":"4.1.0"},"reference-count":26,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2020,4,1]],"date-time":"2020-04-01T00:00:00Z","timestamp":1585699200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["MicrobiologyOpen"],"published-print":{"date-parts":[[2020,5]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Methicillin\u2010resistant <jats:italic>S. aureus<\/jats:italic> has been reported as the main pathogen involved in chronic infections, osteomyelitis, and prosthetic joint infections. The host\/pathogen interaction is dynamic and requires several changes to promote bacterial survival. Here, we focused on the internalization and persistence behavior of well\u2010characterized <jats:italic>Staphylococcus aureus<\/jats:italic> invasive strains belonging to the main ST\u2010MRSA\u2010SCC<jats:italic>mec<\/jats:italic> clones. To overcome the limitations of the cell culture method, we comparatively analyzed the ability of internalization within human MG\u201063 osteoblasts with imaging flow cytometry (IFC). After evaluation by cell culture assay, the MRSA clones in the study were all able to readily internalize at 3h postinfection, the persistence of intracellular bacteria was evaluated at 24h both by routine cell culture and IFC assay, after vancomycin\u2010BODIPY staining. A statistical difference of persistence was found in ST5\u2010SCC<jats:italic>mec<\/jats:italic>II (26.59%), ST228\u2010SCC<jats:italic>mec<\/jats:italic>I (20.25%), ST8\u2010SCC<jats:italic>mec<\/jats:italic>IV (19.52%), ST239\u2010SCC<jats:italic>mec<\/jats:italic>III (47.82%), and ST22\u2010SCC<jats:italic>mec<\/jats:italic>IVh (50.55%) showing the same ability to internalize as ATCC12598 (51%), the invasive isolate used as control strain for invasion and persistence assays. We demonstrated that the intracellular persistence process depends on the total number of infected cells. Comparing our data obtained by IFC with those of the cell culture assay, we obtained greater reproducibility rates and a number of intracellular bacteria, with the advantage of analyzing live host cells. Moreover, with some limitations related to the lack of whole\u2010genome sequencing analysis, we validated the different proclivities to persist in the main Italian HA\u2010MRSA invasive isolates and our results highlighted the heterogeneity of the different clones to persist during cell infection.<\/jats:p>","DOI":"10.1002\/mbo3.1017","type":"journal-article","created":{"date-parts":[[2020,4,1]],"date-time":"2020-04-01T15:34:43Z","timestamp":1585755283000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":19,"title":["Detection of methicillin\u2010resistant <i>Staphylococcus aureus<\/i> persistence in osteoblasts using imaging flow cytometry"],"prefix":"10.1002","volume":"9","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8672-0484","authenticated-orcid":false,"given":"Dafne","family":"Bongiorno","sequence":"first","affiliation":[{"name":"Department of Biomedical and Biotechnological Sciences (BIOMETEC) Medical Molecular Microbiology and Antibiotic Resistance Laboratory (MMAR<i>Lab<\/i>) University of Catania Catania Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2451-1158","authenticated-orcid":false,"given":"Nicol\u00f2","family":"Musso","sequence":"additional","affiliation":[{"name":"Bio\u2010nanotech Research and Innovation Tower (BRIT) University of Catania Catania Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lorenzo Mattia","family":"Lazzaro","sequence":"additional","affiliation":[{"name":"Department of Biomedical and Biotechnological Sciences (BIOMETEC) Medical Molecular Microbiology and Antibiotic Resistance Laboratory (MMAR<i>Lab<\/i>) University of Catania Catania Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0976-2936","authenticated-orcid":false,"given":"Gino","family":"Mongelli","sequence":"additional","affiliation":[{"name":"Department of Biomedical and Biotechnological Sciences (BIOMETEC) Medical Molecular Microbiology and Antibiotic Resistance Laboratory (MMAR<i>Lab<\/i>) University of Catania Catania Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1594-7427","authenticated-orcid":false,"given":"Stefania","family":"Stefani","sequence":"additional","affiliation":[{"name":"Department of Biomedical and Biotechnological Sciences (BIOMETEC) Medical Molecular Microbiology and Antibiotic Resistance Laboratory (MMAR<i>Lab<\/i>) University of Catania Catania Italy"},{"name":"Bio\u2010nanotech Research and Innovation Tower (BRIT) University of Catania Catania Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8405-5425","authenticated-orcid":false,"given":"Floriana","family":"Campanile","sequence":"additional","affiliation":[{"name":"Department of Biomedical and Biotechnological Sciences (BIOMETEC) Medical Molecular Microbiology and Antibiotic Resistance Laboratory (MMAR<i>Lab<\/i>) University of Catania Catania Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2020,4]]},"reference":[{"key":"e_1_2_10_2_1","doi-asserted-by":"publisher","DOI":"10.1089\/mdr.2018.0292"},{"key":"e_1_2_10_3_1","doi-asserted-by":"publisher","DOI":"10.1089\/mdr.2017.0299"},{"key":"e_1_2_10_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jgar.2015.06.006"},{"key":"e_1_2_10_5_1","doi-asserted-by":"publisher","DOI":"10.3390\/ma11040550"},{"key":"e_1_2_10_6_1","first-page":"43","article-title":"Australian Group on Antimicrobial Resistance (AGAR) Australian Staphylococcus aureus Sepsis Outcome Programme (ASSOP) Annual Report 2017","volume":"16","author":"Coombs G. 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