{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,11,12]],"date-time":"2026-11-12T19:02:41Z","timestamp":1794510161920,"version":"4.9.1"},"reference-count":19,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2026,6,19]],"date-time":"2026-06-19T00:00:00Z","timestamp":1781827200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2026,8,31]],"date-time":"2026-08-31T00:00:00Z","timestamp":1788134400000},"content-version":"vor","delay-in-days":73,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["BMC Infect Dis"],"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>Completion of confirmatory HCV RNA testing after anti-HCV reactivity is a critical step in the hepatitis C diagnostic care cascade. However, missed confirmatory testing and repeated anti-HCV testing within routine hospital laboratory workflows remain insufficiently characterized. This study aimed to evaluate the frequency of missed confirmatory HCV RNA testing among anti-HCV-reactive patients, investigate the presence of active infection using archived serum samples, and assess workflow-related diagnostic gaps that could potentially be addressed through laboratory-based reflex testing strategies.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>This retrospective laboratory-based observational study included patients tested for anti-HCV antibodies at a tertiary-care university hospital between January 1 and December 31, 2025. Anti-HCV-reactive patients without confirmatory HCV RNA testing were identified, and retrospective HCV RNA testing was performed on available archived serum samples. Repeated anti-HCV testing patterns and clinic-based differences in confirmatory testing requests were also analyzed to evaluate potential diagnostic workflow gaps.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      Among 27,984 anti-HCV tests performed in 23,833 unique patients, the anti-HCV seroprevalence rate was 1.28%. Confirmatory HCV RNA testing was not requested in 32.3% of anti-HCV-reactive patients. Retrospective testing of archived serum samples from 79 patients without initial confirmatory testing identified active infection in 3.8% of cases (3\/79). Confirmatory testing rates were significantly higher in specialist clinics than in non-specialist clinics (95.0% vs. 71.0%,\n                      <jats:italic>p<\/jats:italic>\n                      \u2009&lt;\u20090.001). Repeated anti-HCV testing was observed in 13.2% of patients, and no seroconversion was detected among individuals tested six or more times.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>Missed confirmatory HCV RNA testing represents an important gap in the diagnostic care cascade, particularly in non-specialist clinical settings. Laboratory-based reflex HCV RNA testing strategies may improve completion of confirmatory testing and reduce unnecessary repeat serological testing, supporting more effective diagnostic stewardship and contributing to hepatitis C elimination efforts.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Clinical trial number<\/jats:title>\n                    <jats:p>Not applicable.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1186\/s12879-026-13807-4","type":"journal-article","created":{"date-parts":[[2026,6,19]],"date-time":"2026-06-19T10:45:14Z","timestamp":1781865914000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Missed confirmatory HCV RNA testing in anti-HCV-reactive patients and the role of reflex testing in improving completion of the hepatitis C diagnostic care cascade: a laboratory-based retrospective study"],"prefix":"10.1186","volume":"26","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1810-8346","authenticated-orcid":false,"given":"\u00d6zg\u00fcr","family":"Appak","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3939-3089","authenticated-orcid":false,"given":"Nilay","family":"Danis","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4228-1213","authenticated-orcid":false,"given":"S\u00fckran","family":"K\u00f6se","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2026,6,19]]},"reference":[{"key":"13807_CR1","unstructured":"World Health Organization. Global progress report on HIV, viral hepatitis and sexually transmitted infections, 2021: accountability for the global health sector strategies 2016\u20132021: actions for impact. Geneva: World Health Organization; 2021."},{"key":"13807_CR2","first-page":"2429","volume":"364","author":"HR Rosen","year":"2011","unstructured":"Rosen HR. Clinical practice. Chronic hepatitis C infection.N Engl. J Med. 2011;364:2429\u201338.","journal-title":"J Med"},{"key":"13807_CR3","unstructured":"T\u00fcrk Karaci\u011fer Ara\u015ft\u0131rmalar\u0131 Derne\u011fi; Viral Hepatitle Sava\u015f\u0131m Derne\u011fi. T\u00fcrkiye hepatit C tan\u0131 ve tedavi k\u0131lavuzu\u20142023. 2023. Accessed December 21, 2024. Available at: https:\/\/www.tkad.org.tr\/wp-content\/uploads\/2023\/11\/TURKIYE-HEPATIT-C-TANI-VE-TEDAVI-KLAVUZU-2023.pdf."},{"issue":"9","key":"13807_CR4","doi-asserted-by":"publisher","first-page":"637","DOI":"10.7326\/M16-2575","volume":"166","author":"O Falade-Nwulia","year":"2017","unstructured":"Falade-Nwulia O, Suarez-Cuervo C, Nelson DR, et al. Oral direct-acting agent therapy for hepatitis C virus infection: a systematic review. Ann Intern Med. 2017;166(9):637\u201348.","journal-title":"Ann Intern Med"},{"issue":"3","key":"13807_CR5","doi-asserted-by":"publisher","first-page":"101626","DOI":"10.1016\/j.clinre.2021.101626","volume":"45","author":"S Roger","year":"2021","unstructured":"Roger S, Ducancelle A, Le Guillou-Guillemette H, Gaudy C, Lunel F. HCV virology and diagnosis. Clin Res Hepatol Gastroenterol. 2021;45(3):101626.","journal-title":"Clin Res Hepatol Gastroenterol"},{"key":"13807_CR6","doi-asserted-by":"publisher","first-page":"48","DOI":"10.1186\/s13036-024-00443-2","volume":"18","author":"AS Baber","year":"2024","unstructured":"Baber AS, Suganthan B, Ramasamy RP. Current advances in hepatitis C diagnostics. J Biol Eng. 2024;18:48.","journal-title":"J Biol Eng"},{"key":"13807_CR7","doi-asserted-by":"publisher","first-page":"1137","DOI":"10.1093\/cid\/ciad126","volume":"77","author":"Y Tao","year":"2023","unstructured":"Tao Y, Tang W, Fajardo E, et al. Reflex hepatitis C virus viral load testing following an initial positive hepatitis C virus antibody test: a global systematic review and meta-analysis. Clin Infect Dis. 2023;77:1137\u201356.","journal-title":"Clin Infect Dis"},{"key":"13807_CR8","doi-asserted-by":"publisher","unstructured":"Manuylov VA, Gushchin VA, Chulanov VP, et al. Regional, age, and sex patterns of hepatitis C virus infection in Russia: insights from a 42,000-participant serosurvey. Viruses. 2025;17:1529. https:\/\/doi.org\/10.3390\/v17121529.","DOI":"10.3390\/v17121529"},{"key":"13807_CR9","doi-asserted-by":"publisher","first-page":"e70087","DOI":"10.1111\/jvh.70087","volume":"32","author":"MG Ghany","year":"2025","unstructured":"Ghany MG, Ward JW, Baldwin Z, et al. HCV testing and treatment of adults in the United States: 2014 through 2021\u2014data from two national commercial testing laboratories. J Viral Hepat. 2025;32:e70087. https:\/\/doi.org\/10.1111\/jvh.70087.","journal-title":"J Viral Hepat"},{"key":"13807_CR10","doi-asserted-by":"publisher","first-page":"1252","DOI":"10.3390\/v17091252","volume":"17","author":"MG Minissale","year":"2025","unstructured":"Minissale MG, Petta S, Cartabellotta F. HCV screening in a Sicilian center: a descriptive cohort. profile Viruses. 2025;17:1252. https:\/\/doi.org\/10.3390\/v17091252.","journal-title":"profile Viruses"},{"key":"13807_CR11","doi-asserted-by":"publisher","first-page":"e01815","DOI":"10.1128\/JCM.01815-19","volume":"58","author":"R L\u00f3pez-Mart\u00ednez","year":"2019","unstructured":"L\u00f3pez-Mart\u00ednez R, Arias-Garc\u00eda A, Rodr\u00edguez-Algarra F, et al. Significant improvement in diagnosis of hepatitis C virus infection by a one-step strategy in a central laboratory: an optimal tool for hepatitis C elimination?J. Clin Microbiol. 2019;58:e01815\u201319.","journal-title":"Clin Microbiol"},{"key":"13807_CR12","doi-asserted-by":"publisher","first-page":"2261","DOI":"10.1111\/liv.14227","volume":"39","author":"SR Bartlett","year":"2019","unstructured":"Bartlett SR, Yu A, Chapinal N, et al. The population level care cascade for hepatitis C in British Columbia, Canada, as of 2018: impact of direct-acting antivirals. Liver Int. 2019;39:2261\u201372.","journal-title":"Liver Int"},{"key":"13807_CR13","unstructured":"World Health Organization. Global health sector strategies on HIV, viral hepatitis and sexually transmitted infections for the period 2022\u20132030. Geneva: World Health Organization; 2022."},{"issue":"Suppl 3","key":"13807_CR14","doi-asserted-by":"publisher","first-page":"S357","DOI":"10.1093\/infdis\/jiad191","volume":"229","author":"LN Johnson","year":"2024","unstructured":"Johnson LN, Gaynor AM, Wroblewski K, Buss SN. Maximizing reflexive hepatitis C virus RNA testing of HCV antibody-reactive samples within United States public health laboratories. J Infect Dis. 2024;229(Suppl 3):S357\u201361.","journal-title":"J Infect Dis"},{"key":"13807_CR15","doi-asserted-by":"publisher","unstructured":"Chen SH, Ding YJ, Hsu NT, et al. Prevalence-guided anti-HCV and reflex HCV antigen testing in the detection of patients with chronic hepatitis C in hepatitis C endemic areas. Diagnostics. 2025;15:3064. https:\/\/doi.org\/10.3390\/diagnostics15233064.","DOI":"10.3390\/diagnostics15233064"},{"key":"13807_CR16","doi-asserted-by":"publisher","unstructured":"Lee MK, Alfego D, Walworth C, Gillim L. Trends in reflexive HCV testing in a national US laboratory: a retrospective study from 2018\u20132024. Clinical Infectious Diseases. 2026:ciag227. https:\/\/doi.org\/10.1093\/cid\/ciag227.","DOI":"10.1093\/cid\/ciag227"},{"key":"13807_CR17","doi-asserted-by":"publisher","first-page":"914","DOI":"10.3855\/jidc.11657","volume":"13","author":"\u00d6 Kiri\u015fci","year":"2019","unstructured":"Kiri\u015fci \u00d6, Cal\u0131skan A. Threshold value of the anti-HCV test in the diagnosis of HCV infection.J Infect. Dev Ctries. 2019;13:914\u20139. https:\/\/doi.org\/10.3855\/jidc.11657.","journal-title":"Dev Ctries"},{"key":"13807_CR18","doi-asserted-by":"publisher","first-page":"484","DOI":"10.18621\/eurj.945588","volume":"9","author":"M Ocal","year":"2023","unstructured":"Ocal M, Bulut ME. Determination of anti-HCV signal-to-cut-off value in patients with hepatitis C virus infection and the variety of antibody responses. Eur Res J. 2023;9:484\u201394. https:\/\/doi.org\/10.18621\/eurj.945588.","journal-title":"Eur Res J"},{"key":"13807_CR19","doi-asserted-by":"publisher","first-page":"e262658","DOI":"10.1001\/jamanetworkopen.2026.2658","volume":"9","author":"N Al Rawashdh","year":"2026","unstructured":"Al Rawashdh N, Ferrufino CP, Kahn EB, et al. Cost-effectiveness of point-of-care hepatitis C virus RNA testing in the United States. JAMA Netw Open. 2026;9:e262658. https:\/\/doi.org\/10.1001\/jamanetworkopen.2026.2658.","journal-title":"JAMA Netw Open"}],"container-title":["BMC Infectious Diseases"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/article\/10.1186\/s12879-026-13807-4","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s12879-026-13807-4.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s12879-026-13807-4.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,31]],"date-time":"2026-08-31T02:11:33Z","timestamp":1788142293000},"score":1,"resource":{"primary":{"URL":"https:\/\/link.springer.com\/10.1186\/s12879-026-13807-4"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,6,19]]},"references-count":19,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2026,12]]}},"alternative-id":["13807"],"URL":"https:\/\/doi.org\/10.1186\/s12879-026-13807-4","relation":{},"ISSN":["1471-2334"],"issn-type":[{"value":"1471-2334","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,6,19]]},"assertion":[{"value":"30 April 2026","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"8 June 2026","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"19 June 2026","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"This study was approved by the Dokuz Eylul University Non-Interventional Research Ethics Committee (approval number: 2025\/02\u201315, date: 15.01.2025) and was conducted in accordance with the Declaration of Helsinki. The requirement for informed consent was waived by the Ethics Committee due to the retrospective design of the study and the use of archived anonymized laboratory data.","order":1,"name":"Ethics","label":"Ethical approval and consent to participate","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"Not applicable.","order":2,"name":"Ethics","label":"Consent for publication","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"The authors declare no competing interests.","order":3,"name":"Ethics","label":"Competing interests","group":{"name":"EthicsHeading","label":"Declarations"}}],"article-number":"1584"}}