{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T19:23:31Z","timestamp":1791401011310,"version":"4.3.3"},"reference-count":19,"publisher":"American Society for Microbiology","issue":"2","license":[{"start":{"date-parts":[[2026,2,11]],"date-time":"2026-02-11T00:00:00Z","timestamp":1770768000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,2,11]],"date-time":"2026-02-11T00:00:00Z","timestamp":1770768000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"DOI":"10.13039\/501100001704","name":"European Society of Clinical Microbiology and Infectious Diseases","doi-asserted-by":"publisher","award":["ESCMIDRG17142"],"award-info":[{"award-number":["ESCMIDRG17142"]}],"id":[{"id":"10.13039\/501100001704","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004587","name":"Instituto de Salud Carlos III","doi-asserted-by":"publisher","award":["CP23CIII\/00004"],"award-info":[{"award-number":["CP23CIII\/00004"]}],"id":[{"id":"10.13039\/501100004587","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004587","name":"Instituto de Salud Carlos III","doi-asserted-by":"publisher","award":["PI19CIII\/00009"],"award-info":[{"award-number":["PI19CIII\/00009"]}],"id":[{"id":"10.13039\/501100004587","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004587","name":"Instituto de Salud Carlos III","doi-asserted-by":"publisher","award":["PI20CIII\/00004"],"award-info":[{"award-number":["PI20CIII\/00004"]}],"id":[{"id":"10.13039\/501100004587","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Centro de Investigaci\u00f3n en Red en Enfermedades Infecciosas","award":["CB21\/13\/00044"],"award-info":[{"award-number":["CB21\/13\/00044"]}]},{"name":"Fundaci\u00f3n para la Investigaci\u00f3n Biom\u00e9dica del Hospital Universitario Pr\u00edncipe de Asturias","award":["VHD-HUPA"],"award-info":[{"award-number":["VHD-HUPA"]}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["J Clin Microbiol"],"published-print":{"date-parts":[[2026,2,11]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title\/>\n                    <jats:p>\n                      Quantitative RT-PCR (qRT-PCR) is essential for monitoring hepatitis delta virus (HDV) RNA, yet assays lack standardization. We aimed to evaluate the performance of three qRT-PCR assays and to assess the impact of a pre-analytical thermal shock procedure. We conducted a comparative study using 206 samples (106 with anti-HDV antibodies and 100 anti-HDV-negative as a control group), which were tested in parallel with three qRT-PCR assays: Vircell, Certest, and Altona. Performance was evaluated for inter-assay agreement (kappa index), quantitative correlation (R\u00b2), and bias (Bland-Altman). Altona detected 56 HDV-RNA positive samples, whereas Vircell and Certest detected 55 positive samples. Inter-assay agreement was perfect comparing Vircell vs Certest (agreement = 100%, \u043a = 1.000) and almost perfect comparing Altona with both Vircell and Certest (agreement = 99.5%, \u043a = 0.988). Quantitatively, Vircell and Certest assays showed relevant systematic biases compared to Altona, overestimating viral loads by approximately 0.24 log\n                      <jats:sub>10<\/jats:sub>\n                      International Units (IU)\/mL (Certest) and 0.33 log\n                      <jats:sub>10<\/jats:sub>\n                      IU\/mL (Vircell). The correlation with Altona was strong for Certest (R\u00b2 = 0.864) and moderate for Vircell (R\u00b2 = 0.793), while the correlation between Certest and Vircell was weaker (R\u00b2 = 0.720). Thermal shock improved sensitivity in one case (Certest vs Vircell) but increased quantitative variability, worsening the inter-assay correlation (R\u00b2 = 0.684). In conclusion, all three assays were highly concordant for the qualitative diagnosis of HDV infection, but their quantitative biases prevent their interchangeable use for treatment monitoring. Thermal shock is not recommended for routine monitoring, as a significant compromise in quantitative accuracy and precision outweighs any potential gains in sensitivity.\n                    <\/jats:p>\n                    <jats:sec>\n                      <jats:title>IMPORTANCE<\/jats:title>\n                      <jats:p>This study evaluates three hepatitis delta virus (HDV) RNA quantitative real-time PCR (qRT-PCR) assays, crucial for managing the HDV infection, particularly in the setting of new therapies like Bulevirtide, where assessing viral load reduction and accurate monitoring is paramount. We reveal significant quantitative biases among widely used assays, precluding their interchangeable use and risking misinterpretation of treatment response. Furthermore, our systematic assessment of the thermal shock pre-analytical procedure highlights its detrimental impact on quantitative precision, despite modest sensitivity gains. This work provides essential evidence for clinicians and laboratories, guiding assay selection and standardization efforts to optimize HDV diagnosis and patient monitoring.<\/jats:p>\n                    <\/jats:sec>\n                  <\/jats:sec>","DOI":"10.1128\/jcm.01517-25","type":"journal-article","created":{"date-parts":[[2025,12,31]],"date-time":"2025-12-31T14:26:36Z","timestamp":1767191196000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":2,"title":["Challenges in accurate HDV RNA quantification: inter-assay variability and the impact of thermal shock"],"prefix":"10.1128","volume":"64","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-4885-4334","authenticated-orcid":false,"given":"Felipe","family":"P\u00e9rez-Garc\u00eda","sequence":"first","affiliation":[{"name":"Servicio de Microbiolog\u00eda Cl\u00ednica, Hospital Universitario Pr\u00edncipe de Asturias","place":["Madrid, Spain"]},{"id":[{"id":"https:\/\/ror.org\/04pmn0e78","id-type":"ROR","asserted-by":"publisher"}],"name":"Universidad de Alcal\u00e1, Facultad de Medicina, Departamento de Biomedicina y Biotecnolog\u00eda","place":["Madrid, Spain"]},{"id":[{"id":"https:\/\/ror.org\/00ca2c886","id-type":"ROR","asserted-by":"publisher"}],"name":"Centro de Investigaci\u00f3n Biom\u00e9dica en Red en Enfermedades Infecciosas (CIBERINFEC), Instituto de Salud Carlos III","place":["Madrid, Spain"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ana","family":"Virseda-Berdices","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/00ca2c886","id-type":"ROR","asserted-by":"publisher"}],"name":"Centro de Investigaci\u00f3n Biom\u00e9dica en Red en Enfermedades Infecciosas (CIBERINFEC), Instituto de Salud Carlos III","place":["Madrid, Spain"]},{"id":[{"id":"https:\/\/ror.org\/00ca2c886","id-type":"ROR","asserted-by":"publisher"}],"name":"Unidad de Infecci\u00f3n Viral e Inmunidad, 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Bernal R Iglesies Torrent J Sellar\u00e8s Crous A Forns X Lens S P\u00e9rez Del Pulgar S Leonel T Hurtado JC. 2024. Evaluaci\u00f3n del rendimiento de una PCR comercial para la detecci\u00f3n de ARN de VHD comparada con una PCR casera. Zaragoza Abstr XXVII Congreso Nacional de la Sociedad Espa\u00f1ola de Enfermedades Infecciosas y Microbiolog\u00eda Cl\u00ednica"}],"container-title":["Journal of Clinical Microbiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/jcm.01517-25","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/jcm.01517-25","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,2,11]],"date-time":"2026-02-11T14:00:58Z","timestamp":1770818458000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/jcm.01517-25"}},"subtitle":[],"editor":[{"given":"Elitza S.","family":"Theel","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2026,2,11]]},"references-count":19,"journal-issue":{"issue":"2","published-print":{"date-parts":[[2026,2,11]]}},"alternative-id":["10.1128\/jcm.01517-25"],"URL":"https:\/\/doi.org\/10.1128\/jcm.01517-25","relation":{},"ISSN":["0095-1137","1098-660X"],"issn-type":[{"value":"0095-1137","type":"print"},{"value":"1098-660X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,2,11]]},"assertion":[{"value":"2025-10-13","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-12-02","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-12-31","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e01517-25"}}