{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,18]],"date-time":"2026-09-18T02:39:18Z","timestamp":1789699158764,"version":"4.0.1"},"reference-count":50,"publisher":"Wiley","issue":"10","license":[{"start":{"date-parts":[[2023,8,13]],"date-time":"2023-08-13T00:00:00Z","timestamp":1691884800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100001691","name":"Japan Society for the Promotion of Science","doi-asserted-by":"publisher","award":["21K07002"],"award-info":[{"award-number":["21K07002"]}],"id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Microbiology and Immunology"],"published-print":{"date-parts":[[2023,10]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>The skin and mucous membranes are the primary sites of <jats:italic>Staphylococcus aureus<\/jats:italic> colonization, particularly those of health care personnel and patients in long\u2010term care centers. We found that <jats:italic>S. aureus<\/jats:italic> colonized with a higher abundance ratio on skins which had recovered from pressure injury (PI) than on normal skins in our earlier research on the skin microbiota of bedridden patients. Multilocus sequence typing (MLST) is a useful tool for typing <jats:italic>S. aureus<\/jats:italic> isolated from clinical specimens. However, the MLST approach cannot be used in microbiota DNA owing to the contamination from other bacteria species. In this study, we developed a multiplex\u2010nested PCR method to determine <jats:italic>S. aureus<\/jats:italic> MLST in samples collected from human skins. The seven pairs of forward and reverse primers were designed in the upstream and downstream regions, which were conserved specifically in <jats:italic>S. aureus<\/jats:italic>. The first amplifications of the seven pairs were conducted in a multiplex assay. The samples were diluted and applied to conventional PCR for MLST. We confirmed that the method amplified the seven allele sequences of <jats:italic>S. aureus<\/jats:italic> specifically in the presence of untargeted DNAs from human and other skin commensal bacteria. Using this assay, we succeeded in typing sequence types (STs) of <jats:italic>S. aureus<\/jats:italic> in the DNA samples derived from the skins healed from PI. Peaks obtained by Sanger sequencing showed that each sample contained one ST, which were mainly categorized into clonal complex 1 (CC1) or CC5. We propose that this culture\u2010free approach may be used in detecting <jats:italic>S. aureus<\/jats:italic> in clinical specimens without isolation.<\/jats:p>","DOI":"10.1111\/1348-0421.13094","type":"journal-article","created":{"date-parts":[[2023,8,14]],"date-time":"2023-08-14T04:40:15Z","timestamp":1691988015000},"page":"438-446","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["A multilocus sequence typing method of <i>Staphylococcus aureus<\/i> DNAs in a sample from human skin"],"prefix":"10.1111","volume":"67","author":[{"given":"Hiroka","family":"Furuya","sequence":"first","affiliation":[{"name":"Department of Clinical Laboratory Science, Faculty of Health Sciences, Institute of Medical, Pharmaceutical and Health Sciences Kanazawa University Kanazawa Ishikawa 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