{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,25]],"date-time":"2026-08-25T18:19:11Z","timestamp":1787681951794,"version":"build-2784847793"},"reference-count":25,"publisher":"Public Library of Science (PLoS)","issue":"8","license":[{"start":{"date-parts":[[2026,8,25]],"date-time":"2026-08-25T00:00:00Z","timestamp":1787616000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["P50CA098252"],"award-info":[{"award-number":["P50CA098252"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["R01CA237067"],"award-info":[{"award-number":["R01CA237067"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["R01CA237067"],"award-info":[{"award-number":["R01CA237067"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["P30CA006973"],"award-info":[{"award-number":["P30CA006973"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS One"],"abstract":"<jats:p>Use of quantitative polymerase chain reaction (qPCR) to measure viral DNA in plasma has promise for the detection and\/or monitoring of high-risk human papillomavirus (hrHPV)-associated cancers. We examined Roche\u2019s cobas 6800 HPV test for detection of hrHPV in plasma or peripheral blood mononuclear cells (PBMC) of patients with HPV-associated dysplasia or cancer. In our pilot study, this assay detected hrHPV DNA in 1 mL pre-treatment plasma of patients with advanced (stage II-IV) cervical cancer, but not stage I or dysplasia cases, or in PBMC. Plasma of oropharyngeal cancer patients also tested positive in most cases. Seeking to enhance sensitivity, we re-tested these pilot samples with qPCR employing three published primer sets (E7, BR E1-5, HR E5L2-4) selected for improved detection of HPV16 in cancer. This triplex assay detected quantifiable levels of HPV16 in pre-treatment plasma of advanced (stage II-IV) cervical cancer cases, and lower levels in some stage I and dysplasia cases. In serial sampling of HPV16\u2009+\u2009cervical cancer patients post-treatment with no evidence of disease over ~2 years, no reproducible signal was detected by cobas HPV testing, whereas the triplex qPCR sporadically detected HPV DNA in plasma and PBMC samples of a subset of these patients.<\/jats:p>","DOI":"10.1371\/journal.pone.0354671","type":"journal-article","created":{"date-parts":[[2026,8,25]],"date-time":"2026-08-25T17:27:24Z","timestamp":1787678844000},"page":"e0354671","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":0,"title":["Application of Cobas 6800 HPV testing and qPCR utilizing three genomically-optimized primer sets for detecting HPV DNA in the blood of HPV Positive dysplasia and cancer patients"],"prefix":"10.1371","volume":"21","author":[{"given":"Heba H.","family":"Mostafa","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Amary","family":"Fall","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"St\u00e9phanie","family":"Gaillard","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carole","family":"Fakhry","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zubair","family":"Khan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tanguy","family":"Seiwert","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hao","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8388-6297","authenticated-orcid":true,"given":"Hua-Ling","family":"Tsai","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Amanda N.","family":"Fader","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4034-6845","authenticated-orcid":true,"given":"Charles A.","family":"Leath","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Warner K.","family":"Huh","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rebecca C.","family":"Arend","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ya-chea","family":"Tsai","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joseph A.","family":"Califano","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Deyin","family":"Xing","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"T.-C.","family":"Wu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2506-0195","authenticated-orcid":true,"given":"Richard B. S.","family":"Roden","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2026,8,25]]},"reference":[{"issue":"5","key":"pone.0354671.ref001","doi-asserted-by":"crossref","first-page":"342","DOI":"10.1038\/nrc798","article-title":"Papillomaviruses and cancer: from basic studies to clinical application","volume":"2","author":"H zur Hausen","year":"2002","journal-title":"Nat Rev Cancer"},{"issue":"7","key":"pone.0354671.ref002","doi-asserted-by":"crossref","first-page":"1336","DOI":"10.1002\/ijc.35278","article-title":"The GLOBOCAN 2022 cancer estimates: Data sources, methods, and a snapshot of the cancer burden worldwide","volume":"156","author":"AM Filho","year":"2025","journal-title":"Int J Cancer"},{"issue":"8","key":"pone.0354671.ref003","article-title":"Cytology versus HPV testing for cervical cancer screening in the general population","volume":"8","author":"G Koliopoulos","year":"2017","journal-title":"Cochrane Database Syst Rev"},{"issue":"6","key":"pone.0354671.ref004","doi-asserted-by":"crossref","first-page":"547","DOI":"10.1001\/jama.2023.13174","article-title":"Cervical Cancer Screening: A Review","volume":"330","author":"RB Perkins","year":"2023","journal-title":"JAMA"},{"issue":"5","key":"pone.0354671.ref005","doi-asserted-by":"crossref","first-page":"157","DOI":"10.1080\/14737159.2025.2486653","article-title":"Evolution and future of cervical cancer screening: from cytology to primary HPV testing and the impact of vaccination","volume":"25","author":"M El-Zein","year":"2025","journal-title":"Expert Rev Mol Diagn"},{"issue":"2","key":"pone.0354671.ref006","doi-asserted-by":"crossref","DOI":"10.1016\/j.ajog.2016.02.021","article-title":"Impact of human papillomavirus (HPV) 16 and 18 vaccination on prevalent infections and rates of cervical lesions after excisional treatment","volume":"215","author":"A Hildesheim","year":"2016","journal-title":"Am J Obstet Gynecol"},{"issue":"14","key":"pone.0354671.ref007","doi-asserted-by":"crossref","first-page":"2541","DOI":"10.3390\/cancers16142541","article-title":"Management for Cervical Cancer Patients: A Comparison of the Guidelines from the International Scientific Societies (ESGO-NCCN-ASCO-AIOM-FIGO-BGCS-SEOM-ESMO-JSGO)","volume":"16","author":"S Restaino","year":"2024","journal-title":"Cancers (Basel)"},{"key":"pone.0354671.ref008","doi-asserted-by":"crossref","first-page":"105805","DOI":"10.1016\/j.oraloncology.2022.105805","article-title":"Comparison of next generation sequencing, droplet digital PCR, and quantitative real-time PCR for the earlier detection and quantification of HPV in HPV-positive oropharyngeal cancer","volume":"128","author":"AK Mattox","year":"2022","journal-title":"Oral Oncol"},{"issue":"11","key":"pone.0354671.ref009","doi-asserted-by":"crossref","first-page":"2097","DOI":"10.1038\/s41416-023-02233-x","article-title":"Detection of circulating cell-free HPV DNA of 13 HPV types for patients with cervical cancer as potential biomarker to monitor therapy response and to detect relapse","volume":"128","author":"S Mittelstadt","year":"2023","journal-title":"Br J Cancer"},{"key":"pone.0354671.ref010","article-title":"Direct comparison of alternative blood-based approaches for early detection and diagnosis of HPV-associated head and neck cancers","author":"ME Bryan","year":"2025","journal-title":"Clin Cancer Res"},{"issue":"4","key":"pone.0354671.ref011","doi-asserted-by":"crossref","first-page":"725","DOI":"10.3390\/diagnostics13040725","article-title":"Analytical Validation of NavDx, a cfDNA-Based Fragmentomic Profiling Assay for HPV-Driven Cancers","volume":"13","author":"A Gunning","year":"2023","journal-title":"Diagnostics (Basel)"},{"issue":"15","key":"pone.0354671.ref012","doi-asserted-by":"crossref","first-page":"3150","DOI":"10.1158\/1078-0432.CCR-25-0152","article-title":"Early Dynamics of Circulating Tumor HPV-DNA with Neoadjuvant Chemotherapy and Response-Adapted De-escalation in Human Papillomavirus-Associated Oropharyngeal Cancer","volume":"31","author":"AJ Rosenberg","year":"2025","journal-title":"Clin Cancer Res"},{"issue":"5","key":"pone.0354671.ref013","doi-asserted-by":"crossref","first-page":"488","DOI":"10.1001\/jamaoto.2022.0159","article-title":"Association of Plasma Circulating Tumor HPV DNA With HPV-Related Oropharynx Cancer","volume":"148","author":"SR Tewari","year":"2022","journal-title":"JAMA Otolaryngol Head Neck Surg"},{"issue":"5","key":"pone.0354671.ref014","doi-asserted-by":"crossref","first-page":"1478","DOI":"10.1128\/JCM.03386-12","article-title":"Development and characterization of the cobas human papillomavirus test","volume":"51","author":"A Rao","year":"2013","journal-title":"J Clin Microbiol"},{"issue":"4","key":"pone.0354671.ref015","doi-asserted-by":"crossref","first-page":"688","DOI":"10.1002\/hed.26041","article-title":"Rational genomic optimization of DNA detection for human papillomavirus type 16 in head and neck squamous cell carcinoma","volume":"42","author":"Y Saito","year":"2020","journal-title":"Head Neck"},{"key":"pone.0354671.ref016","doi-asserted-by":"crossref","first-page":"86","DOI":"10.1016\/j.ygyno.2025.08.006","article-title":"A randomized pilot study of HPV16 L2E7E6 fusion protein vaccination site post-treatment for HPV16+ cervical cancer","volume":"201","author":"S Gaillard","year":"2025","journal-title":"Gynecol Oncol"},{"issue":"15","key":"pone.0354671.ref017","doi-asserted-by":"crossref","first-page":"3329","DOI":"10.1158\/1078-0432.CCR-24-0590","article-title":"Personalized ctDNA for Monitoring Disease Status in Head and Neck Squamous Cell Carcinoma","volume":"30","author":"GJ Hanna","year":"2024","journal-title":"Clinical Cancer Research"},{"issue":"9","key":"pone.0354671.ref018","doi-asserted-by":"crossref","first-page":"1605","DOI":"10.1158\/1078-0432.CCR-24-3053","article-title":"A Prospective Trial of Biomarker-Guided Surveillance for HPV-Positive Oropharynx Cancer Using Plasma Tumor Tissue-Modified Viral HPV DNA","volume":"31","author":"EM Rettig","year":"2025","journal-title":"Clin Cancer Res"},{"issue":"9","key":"pone.0354671.ref019","doi-asserted-by":"crossref","first-page":"1561","DOI":"10.1158\/1078-0432.CCR-25-0012","article-title":"So close, and yet. plasma HPV DNA detection of recurrent HPV oropharynx cancer","volume":"31","author":"JA Califano","year":"2025","journal-title":"Clin Cancer Res"},{"issue":"4","key":"pone.0354671.ref020","first-page":"263","article-title":"Human papilloma virus detection in oropharyngeal cancer with gargle samples","volume":"12","author":"T Imai","year":"2016","journal-title":"B-ENT"},{"issue":"6","key":"pone.0354671.ref021","doi-asserted-by":"crossref","first-page":"408","DOI":"10.1136\/sextrans-2019-054359","article-title":"Identification of human papillomavirus in oral rinse specimens from women with and without cervical intraepithelial lesions","volume":"96","author":"I Nemesio","year":"2020","journal-title":"Sex Transm Infect"},{"issue":"7","key":"pone.0354671.ref022","doi-asserted-by":"crossref","first-page":"692","DOI":"10.1111\/jop.13172","article-title":"Prevalence and concordance of oral HPV infections with cervical HPV infections in women referred to colposcopy with abnormal cytology","volume":"50","author":"P Tewari","year":"2021","journal-title":"J Oral Pathol Med"},{"key":"pone.0354671.ref023","doi-asserted-by":"crossref","first-page":"107262","DOI":"10.1016\/j.oraloncology.2025.107262","article-title":"Circulating tumor DNA in human papillomavirus-associated oropharyngeal cancer management: A systematic review","volume":"164","author":"S Chennareddy","year":"2025","journal-title":"Oral Oncol"},{"key":"pone.0354671.ref024","doi-asserted-by":"crossref","first-page":"107249","DOI":"10.1016\/j.oraloncology.2025.107249","article-title":"Oral HPV incidence and risk factors for acquisition","volume":"163","author":"G DSouza","year":"2025","journal-title":"Oral Oncol"},{"issue":"4","key":"pone.0354671.ref025","doi-asserted-by":"crossref","first-page":"516","DOI":"10.1158\/1055-9965.EPI-23-1272","article-title":"Oncogenic Oral Human Papillomavirus Clearance Patterns over 10 Years","volume":"33","author":"G D\u2019Souza","year":"2024","journal-title":"Cancer Epidemiology, Biomarkers & Prevention"}],"container-title":["PLOS One"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0354671","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,25]],"date-time":"2026-08-25T17:27:29Z","timestamp":1787678849000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0354671"}},"subtitle":[],"editor":[{"given":"Maria Lina","family":"Tornesello","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2026,8,25]]},"references-count":25,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2026,8,25]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0354671","relation":{},"ISSN":["1932-6203"],"issn-type":[{"value":"1932-6203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,25]]}}}