{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T06:12:19Z","timestamp":1788415939898,"version":"build-2803163510"},"reference-count":24,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T00:00:00Z","timestamp":1788393600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100018688","name":"Korea Disease Control and Prevention Agency","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100018688","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100013128","name":"Korea Technology and Information Promotion Agency for SMEs","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100013128","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Cell. Infect. Microbiol."],"abstract":"<jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>\n                      Non-invasive molecular tools that can both screen for severe periodontitis and quantify treatment-associated pathogen changes remain an unmet need. We evaluated a non-invasive multiplex quantitative polymerase chain reaction (qPCR) mouthwash panel providing absolute colony-forming-unit (CFU) quantification of five periodontal pathogens (\n                      <jats:italic>Porphyromonas gingivalis, Treponema denticola, Tannerella forsythia, Prevotella intermedia, Fusobacterium nucleatum<\/jats:italic>\n                      ), and asked whether a single test can both screen for severe disease and track each patient\u2019s species-level treatment response.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>Treatment-response monitoring was assessed in a real-world retrospective cohort (349 patients, 703 visits; 235 paired pre\/post-treatment samples) using paired Wilcoxon signed-rank tests and Jonckheere\u2013Terpstra trend tests across baseline stage. Screening performance was independently evaluated in a prospective cross-sectional cohort (n=87; 31 healthy, 56 Stage III\/IV) with leave-one-out cross-validation (LOOCV) and a pre-specified sensitivity analysis.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      For screening, LOOCV AUC was 0.96 for\n                      <jats:italic>P. gingivalis<\/jats:italic>\n                      and 0.97 for the combined panel, preserved under inclusion of antibiotic- and treatment-exposed subjects. Beyond screening, the panel resolved each patient\u2019s species-level response to non-surgical therapy: all five species decreased significantly, with median within-patient reductions of \u22120.37 to \u22120.72 log\n                      <jats:sub>10<\/jats:sub>\n                      CFU (paired Wilcoxon, all p&amp;lt;0.001) and reduction magnitude scaling with baseline severity. This response was strongly species-specific and clinically informative at the individual level: In the implant-bearing high-baseline-burden subgroup, post-treatment\n                      <jats:italic>F. nucleatum<\/jats:italic>\n                      reached or fell below the Healthy reference median in 61% of evaluable patients, whereas\n                      <jats:italic>P. gingivalis<\/jats:italic>\n                      reached it in only 5% and remained approximately 500-fold above the Healthy reference at the median final visit, identifying residual keystone-pathogen burden not captured by clinical or radiographic assessment.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>\n                      In this two-cohort evaluation, a single non-invasive mouthwash qPCR test provided two linked clinical outputs: high-accuracy screening for severe periodontitis and quantitative individual-level tracking of species-specific microbial response to therapy. These findings are preliminary, derive from a single-center screening cohort and a retrospective monitoring cohort, and require external validation before clinical deployment. By placing pathogen loads on the same absolute CFU scale, the assay supports both cross-sectional disease detection and longitudinal residual-burden monitoring as an adjunct to the 2018 AAP\/EFP framework. Clinical Research Information Service (CRIS) registration: KCT0011511 (\n                      <jats:ext-link>https:\/\/cris.nih.go.kr<\/jats:ext-link>\n                      ).\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fcimb.2026.1917870","type":"journal-article","created":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T05:13:54Z","timestamp":1788412434000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["Absolute quantification by mouthwash testing screens severe periodontitis and tracks species-specific microbial response to periodontal therapy"],"prefix":"10.3389","volume":"16","author":[{"given":"Je-Hyun","family":"Eom","sequence":"first","affiliation":[{"name":"Apple Tree Institute of Biomedical Science, Apple Tree Medical Foundation","place":["Goyang, Republic of Korea"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mu-Yeol","family":"Cho","sequence":"additional","affiliation":[{"name":"Apple Tree Institute of Biomedical Science, Apple Tree Medical Foundation","place":["Goyang, Republic of Korea"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ji-Won","family":"Kim","sequence":"additional","affiliation":[{"name":"Apple Tree Institute of Biomedical Science, Apple Tree Medical Foundation","place":["Goyang, Republic of Korea"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yunwoo","family":"Kim","sequence":"additional","affiliation":[{"name":"Apple Tree Institute of Biomedical Science, Apple Tree Medical Foundation","place":["Goyang, Republic of Korea"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jeehyun","family":"No","sequence":"additional","affiliation":[{"name":"Apple Tree Dental Hospital, Apple Tree Medical Foundation","place":["Goyang, Republic of Korea"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiyoung","family":"Hwang","sequence":"additional","affiliation":[{"name":"Research and Development Center, DOCSMEDI Co. Ltd.","place":["Goyang, Republic of Korea"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Seokwon","family":"Kim","sequence":"additional","affiliation":[{"name":"Research and Development Center, DOCSMEDI Co. Ltd.","place":["Goyang, Republic of Korea"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dahye","family":"Lee","sequence":"additional","affiliation":[{"name":"Apple Tree Dental Hospital, Apple Tree Medical Foundation","place":["Goyang, Republic of Korea"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hanseung","family":"Baek","sequence":"additional","affiliation":[{"name":"Apple Tree Dental Hospital, Apple Tree Medical Foundation","place":["Goyang, Republic of Korea"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bae-Kyung","family":"Kim","sequence":"additional","affiliation":[{"name":"Since 1982 THE Lee Hae-seung Dental Clinic","place":["Paju-si, Gyeonggi-do, Republic of Korea"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Young-Youn","family":"Kim","sequence":"additional","affiliation":[{"name":"Apple Tree Dental Hospital, Apple Tree Medical Foundation","place":["Goyang, Republic of Korea"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Soo-Bok","family":"Her","sequence":"additional","affiliation":[{"name":"Artificial Intelligence Research Center, Digital Dental Hub Incorporation","place":["Seoul, Republic of Korea"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dong-Yub","family":"Ko","sequence":"additional","affiliation":[{"name":"Artificial Intelligence Research Center, Digital Dental Hub Incorporation","place":["Seoul, Republic of Korea"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Inseong","family":"Hwang","sequence":"additional","affiliation":[{"name":"COREE Pohang Co., Ltd.","place":["Pohang-si, Gyeongsangbuk-do, Republic of Korea"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hye-Sung","family":"Kim","sequence":"additional","affiliation":[{"name":"Apple Tree Dental Hospital, Apple Tree Medical Foundation","place":["Goyang, Republic of Korea"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,9,3]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0182992","article-title":"Microbial profile comparisons of saliva, pooled and site-specific subgingival samples in periodontitis patients","volume":"12","author":"Belstr\u00f8m","year":"2017","journal-title":"PloS One"},{"key":"B2","doi-asserted-by":"publisher","first-page":"h5527","DOI":"10.1136\/bmj.h5527","article-title":"STARD 2015: an updated list of essential items for reporting diagnostic accuracy studies","volume":"351","author":"Bossuyt","year":"2015","journal-title":"BMJ"},{"key":"B3","doi-asserted-by":"crossref","DOI":"10.1373\/clinchem.2008.112797","article-title":"The MIQE guidelines: minimum information forpublication of quantitative real-time PCR experiments","volume-title":"Clin. Chem.","author":"Bustin","year":"2009"},{"key":"B4","doi-asserted-by":"publisher","first-page":"634","DOI":"10.1093\/clinchem\/hvaf043","article-title":"MIQE 2.0: revision of the minimum information for publication of quantitative real-time PCR experiments guidelines","volume":"71","author":"Bustin","year":"2025","journal-title":"Clin. Chem."},{"key":"B5","doi-asserted-by":"publisher","first-page":"7531","DOI":"10.1038\/s41598-020-64509-z","article-title":"Deep learning hybrid method to automatically diagnose periodontal bone loss and stage periodontitis","volume":"10","author":"Chang","year":"2020","journal-title":"Sci. Rep."},{"key":"B6","doi-asserted-by":"publisher","DOI":"10.3389\/fcimb.2026.1809787","article-title":"Oral microbiome profiles by periodontitis stage in a Korean population","volume":"16","author":"Cho","year":"2026","journal-title":"Front. Cell. Infect. Microbiol."},{"key":"B7","doi-asserted-by":"publisher","first-page":"717","DOI":"10.1038\/nrmicro2873","article-title":"The keystone-pathogen hypothesis","volume":"10","author":"Curtis","year":"2012","journal-title":"Nat. Rev. Microbiol."},{"key":"B8","doi-asserted-by":"publisher","first-page":"748","DOI":"10.1111\/jre.13377","article-title":"Diagnostic accuracy of microbiome\u2010derived biomarkers in periodontitis: Systematic review and meta\u2010analysis","volume":"60","author":"Dong","year":"2025","journal-title":"J. Periodontal Res."},{"key":"B9","doi-asserted-by":"publisher","DOI":"10.1080\/20002297.2022.2145729","article-title":"Fusobacterium nucleatum and its associated systemic diseases: Epidemiologic studies and possible mechanisms","volume":"15","author":"Fan","year":"2023","journal-title":"J. Oral. Microbiol."},{"key":"B10","doi-asserted-by":"publisher","first-page":"261","DOI":"10.1902\/jop.1961.32.3.261","article-title":"Dimensions and relations of the dentogingival junction in humans","volume":"32","author":"Gargiulo","year":"1961","journal-title":"J. Periodontology"},{"key":"B11","doi-asserted-by":"publisher","first-page":"409","DOI":"10.1111\/j.2041-1014.2012.00663.x","article-title":"Beyond the red complex and into more complexity: The polymicrobial synergy and dysbiosis (PSD) model of periodontal disease etiology","volume":"27","author":"Hajishengallis","year":"2012","journal-title":"Mol. Oral. Microbiol."},{"key":"B12","doi-asserted-by":"publisher","first-page":"528","DOI":"10.1016\/j.chom.2023.02.009","article-title":"Oral polymicrobial communities: Assembly, function, and impact on diseases","volume":"31","author":"Hajishengallis","year":"2023","journal-title":"Cell. Host Microbe"},{"key":"B13","doi-asserted-by":"publisher","first-page":"444","DOI":"10.5051\/jpis.2304520226","article-title":"Highly accurate measurement of the relative abundance of oral pathogenic bacteria using colony-forming unit-based qPCR","volume":"54","author":"Hwang","year":"2024","journal-title":"J. Periodontal Implant Sci."},{"key":"B14","doi-asserted-by":"publisher","DOI":"10.3390\/antibiotics13060571","article-title":"Supra-and subgingival microbiome in gingivitis and impact of biofilm control: A comprehensive review","volume":"13","author":"Iniesta","year":"2024","journal-title":"Antibiotics"},{"key":"B15","doi-asserted-by":"publisher","first-page":"394","DOI":"10.5051\/jpis.2200680034","article-title":"Comparison of the oral microbial composition between healthy individuals and periodontitis patients in different oral sampling sites using 16S metagenome profiling","volume":"52","author":"Kim","year":"2022","journal-title":"J. Periodontal Implant Sci."},{"key":"B16","doi-asserted-by":"publisher","first-page":"963","DOI":"10.1111\/jre.13409","article-title":"The effect of non\u2010surgical periodontal therapy on subgingival microbiota: A systematic review and meta\u2010analysis","volume":"60","author":"Krajewski","year":"2025","journal-title":"J. Periodontal Res."},{"key":"B17","doi-asserted-by":"publisher","first-page":"85","DOI":"10.1111\/prd.12147","article-title":"Microbial colonization of the periodontal pocket and its significance for periodontal therapy","volume":"76","author":"Mombelli","year":"2018","journal-title":"Periodontology 2000"},{"key":"B18","doi-asserted-by":"publisher","first-page":"S173","DOI":"10.1111\/jcpe.12946","article-title":"Periodontitis: Consensus report of workgroup 2 of the 2017 World Workshop on the Classification of Periodontal and Peri\u2010Implant Diseases and Conditions","volume":"89","author":"Papapanou","year":"2018","journal-title":"J. Periodontology"},{"key":"B19","doi-asserted-by":"publisher","first-page":"134","DOI":"10.1111\/j.1600-051x.1998.tb02419.x","article-title":"Microbial complexes in subgingival plaque","volume":"25","author":"Socransky","year":"1998","journal-title":"J. Clin. Periodontology"},{"key":"B20","doi-asserted-by":"publisher","first-page":"128","DOI":"10.1097\/ede.0b013e3181c30fb2","article-title":"Assessing the performance of prediction models: A framework for traditional and novel measures","volume":"21","author":"Steyerberg","year":"2010","journal-title":"Epidemiology"},{"key":"B21","doi-asserted-by":"publisher","first-page":"S159","DOI":"10.1111\/jcpe.12945","article-title":"Staging and grading of periodontitis: Framework and proposal of a new classification and case definition","volume":"89","author":"Tonetti","year":"2018","journal-title":"J. Periodontology"},{"key":"B22","doi-asserted-by":"publisher","first-page":"i6","DOI":"10.1136\/bmj.i6","article-title":"Net benefit approaches to the evaluation of prediction models, molecular markers, and diagnostic tests","volume":"352","author":"Vickers","year":"2016","journal-title":"BMJ"},{"key":"B23","doi-asserted-by":"publisher","first-page":"1453","DOI":"10.1136\/bmj.39335.541782.ad","article-title":"The Strengthening the Reporting of Observational Studies in Epidemiology (STROBE) statement: Guidelines for reporting observational studies","volume":"370","author":"Von Elm","year":"2007","journal-title":"Lancet"},{"key":"B24","doi-asserted-by":"publisher","first-page":"861","DOI":"10.1186\/s12903-025-06271-0","article-title":"Burden of severe periodontitis: New insights based on a systematic analysis from the Global Burden of Disease Study 2021","volume":"25","author":"Wu","year":"2025","journal-title":"BMC Oral. Health"}],"container-title":["Frontiers in Cellular and Infection Microbiology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fcimb.2026.1917870\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T05:13:56Z","timestamp":1788412436000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fcimb.2026.1917870\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,3]]},"references-count":24,"alternative-id":["10.3389\/fcimb.2026.1917870"],"URL":"https:\/\/doi.org\/10.3389\/fcimb.2026.1917870","relation":{},"ISSN":["2235-2988"],"issn-type":[{"value":"2235-2988","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,3]]},"article-number":"1917870"}}