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Samples were reduced, alkylated, and trypsinized overnight, followed by analysis using liquid chromatography\u2010tandem mass spectrometry (LC\u2010MS\/MS) on a high\u2010resolution Orbitrap Eclipse mass spectrometer. MaxQuant software suite was used for protein identification, filtering, and label\u2010free quantitation.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Principal component analysis of the identified proteins clearly separated <jats:italic>S. aureus<\/jats:italic> PJI and NIAF samples. Overall, 810 proteins were identified based on their detection in at least three out of four samples from each group; 35 statistically significant differentially abundant proteins (DAPs) were found (two\u2010sample <jats:italic>t<\/jats:italic>\u2010test <jats:italic>p<\/jats:italic>\u2010values \u22640.05 and log<jats:sub>2<\/jats:sub>fold\u2010change values \u22652 or \u2264\u22122). Gene ontology pathway analysis found that microbial defense responses, specifically those related to neutrophil activation, to be increased in <jats:italic>S. aureus<\/jats:italic> PJI compared to NIAF samples.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion and clinical relevance<\/jats:title><jats:p>Proteomic profiling of sonicate fluid using LC\u2010MS\/MS differentiated <jats:italic>S. aureus<\/jats:italic> PJI and NIAF in this pilot study. Further work is needed using a larger sample size and including non\u2010<jats:italic>S. aureus<\/jats:italic> PJI and a diversty of NIAF\u2010types.<\/jats:p><\/jats:sec>","DOI":"10.1002\/prca.202200071","type":"journal-article","created":{"date-parts":[[2023,3,20]],"date-time":"2023-03-20T10:05:59Z","timestamp":1679306759000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Mass spectrometry\u2010based proteomic profiling of sonicate fluid differentiates <i>Staphylococcus aureus<\/i> periprosthetic joint infection from non\u2010infectious failure: A pilot study"],"prefix":"10.1002","volume":"17","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7230-1078","authenticated-orcid":false,"given":"Cody R.","family":"Fisher","sequence":"first","affiliation":[{"name":"Mayo Clinic Graduate School of Biomedical Sciences, Department of Immunology Mayo Clinic  Rochester Minnesota USA"},{"name":"Division of Clinical Microbiology, Department of Laboratory Medicine and Pathology Mayo Clinic  Rochester Minnesota USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5791-727X","authenticated-orcid":false,"given":"Kiran K.","family":"Mangalaparthi","sequence":"additional","affiliation":[{"name":"Division of Clinical Biochemistry and Immunology, Department of Laboratory Medicine and Pathology Mayo Clinic  Rochester Minnesota USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3760-4310","authenticated-orcid":false,"given":"Kerryl E.","family":"Greenwood\u2010Quaintance","sequence":"additional","affiliation":[{"name":"Division of Clinical Microbiology, Department of Laboratory Medicine and Pathology Mayo Clinic  Rochester Minnesota USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2398-1724","authenticated-orcid":false,"given":"Matthew P.","family":"Abdel","sequence":"additional","affiliation":[{"name":"Department of Orthopedic Surgery Mayo Clinic  Rochester Minnesota USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9943-6127","authenticated-orcid":false,"given":"Akhilesh","family":"Pandey","sequence":"additional","affiliation":[{"name":"Division of Clinical Biochemistry and Immunology, Department of Laboratory Medicine and Pathology Mayo Clinic  Rochester Minnesota USA"},{"name":"Manipal Academy of Higher Education  Manipal Karnataka India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6344-4141","authenticated-orcid":false,"given":"Robin","family":"Patel","sequence":"additional","affiliation":[{"name":"Division of Clinical Microbiology, Department of Laboratory Medicine and Pathology Mayo Clinic  Rochester Minnesota USA"},{"name":"Division of Public Health, Infectious Diseases and Occupational Medicine, Department of Medicine Mayo Clinic  Rochester Minnesota USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2023,3,27]]},"reference":[{"key":"e_1_2_11_2_1","doi-asserted-by":"publisher","DOI":"10.1128\/CMR.00111\u201013"},{"key":"e_1_2_11_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.berh.2006.08.003"},{"key":"e_1_2_11_4_1","doi-asserted-by":"publisher","DOI":"10.1097\/01.qco.0000235161.85925.e8"},{"key":"e_1_2_11_5_1","doi-asserted-by":"publisher","DOI":"10.2106\/JBJS.N.01141"},{"key":"e_1_2_11_6_1","doi-asserted-by":"publisher","DOI":"10.1056\/nejmra2203477"},{"key":"e_1_2_11_7_1","doi-asserted-by":"publisher","DOI":"10.2106\/JBJS.F.00222"},{"key":"e_1_2_11_8_1","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1007\/978-3-319-30091-7_2","volume-title":"The incidence and socioeconomic Impact of periprosthetic joint infection: United States perspective Periprosthetic Joint Infections","author":"Gutowski C. 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