485675d3d2841b7eb9beb255a7451d65 pmc_9045303.pdf 3bed5ecc072c20c6c654c507a5f3558679465b41 pmc_9045303.pdf 8709188a72377b9ffc0b6c39f7c7c5d1e3f85d63106b7983a157a0aad061a2b7 pmc_9045303.pdf Title: Standardized Hepatitis B Virus RNA Quantification in Untreated and Treated Chronic Patients: a Promising Marker of Infection Follow-Up Subject: Research Article Keywords: HBV; HBV-RNA; RT-PCR; antiviral treatment; biomonitoring Author: Maria Francesca Cortese;Mar Riveiro-Barciela;David Tabernero;Francisco Rodriguez-Algarra;Adriana Palom;Sara Sopena;Ariadna Rando-Segura;Luisa Roade;Alison Kuchta;Roser Ferrer-Costa;Josep Quer;Beatriz Pacin;Marta Vila;Rosario Casillas;Selene Garcia-Garcia;Rafael Esteban;Tomás Pumarola;Maria Buti;Francisco Rodriguez-Frias Creator: PTC Arbortext Advanced Print Publisher 11.1.4012/W Producer: Adobe LiveCycle PDF Generator; modified using iText® 5.5.13.2 ©2000-2020 iText Group NV (AGPL-version) CreationDate: Fri Apr 15 17:25:07 2022 CEST ModDate: Sat Apr 16 22:18:59 2022 CEST Custom Metadata: no Metadata Stream: yes Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 9 Encrypted: no Page size: 584.957 x 782.986 pts Page rot: 0 File size: 623628 bytes Optimized: no PDF version: 1.4 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- JGDEDJ+AdvOTce3d9a73 Type 1C WinAnsi yes yes no 86 0 JGDEDK+AdvOTce3d9a73+fb Type 1C Custom yes yes yes 313 0 JGDEEL+AdvOT07517017 Type 1C WinAnsi yes yes no 314 0 JGDEEM+AdvOT3b30f6db.B Type 1C WinAnsi yes yes no 36 0 JGDEEN+AdvOT46dcae81 Type 1C WinAnsi yes yes no 37 0 JGDEEO+AdvOT46dcae81+fb Type 1C Custom yes yes yes 38 0 JGDEFP+AdvOT65f8a23b.I Type 1C WinAnsi yes yes no 54 0 JGDEGA+AdvOT8817665d Type 1C WinAnsi yes yes no 39 0 JGDEGB+AdvOT07517017+fb Type 1C Custom yes yes yes 315 0 JGDEIB+AdvP80675 Type 1C WinAnsi yes yes no 40 0 JGDEIC+AdvOT46dcae81+20 Type 1C WinAnsi yes yes no 41 0 JGDFAA+Arial-BoldMT Type 1C WinAnsi yes yes no 67 0 JGDFDP+AdvOT56309c18.I Type 1C WinAnsi yes yes no 83 0 JGDFFA+AdvOT3b30f6db.B+fb Type 1C Custom yes yes yes 132 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2024-10-12 02:44:47 CEST RepresentationInformation: pmc_9045303.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2024-10-01 02:43:00 CEST Size: 623628 Format: PDF Version: 1.4 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 355 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: ViewerPreferences: HideToolbar: false HideMenubar: false HideWindowUI: false FitWindow: false CenterWindow: false DisplayDocTitle: true NonFullScreenPageMode: UseNone Direction: L2R ViewArea: CropBox ViewClip: CropBox PrintArea: CropBox PageClip: CropBox PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: https://doi.org/ Standardized Hepatitis B Virus RNA Quantification in Untreated and Treated Chronic Patients: a Promising Marker of Infection Follow-Up 10.1128/spectrum.02149-21 Maria Francesca Cortese Mar Riveiro-Barciela David Tabernero Francisco Rodriguez-Algarra Adriana Palom Sara Sopena Ariadna Rando-Segura Luisa Roade Alison Kuchta Roser Ferrer-Costa Josep Quer Beatriz Pacin Marta Vila Rosario Casillas Selene Garcia-Garcia Rafael Esteban Tomás Pumarola Maria Buti Francisco Rodriguez-Frias Research Article Research Article HBV HBV-RNA RT-PCR antiviral treatment biomonitoring American Society for Microbiology Cortese et al. Copyright © 2022 Cortese et al. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license. 20220404 2022 ABSTRACT The measurement and interpretation of HBV DNA and RNA levels in HBV infected patients treated with antiviral therapy supports the objective of HBV disease management. Here, we quantified circulating HBV RNA through a standardized and sensitive assay in follow-up samples from both naive and treated patients as a marker of infection evolution. HBV DNA (HBV DNA for use in Cobas 6800/8800 Automated Roche Molecular Systems), RNA (Roche HBV RNA Investigational Assay for use in the Cobas 6800/8800; Roche), HBeAg and HBsAg (Elycsys HBsAg chemiluminescence immunoassay by Cobas 8000; Roche), and core-related antigen (Lumipulse G chemiluminescence assay; Fujirebio) levels were measured in cohorts of untreated or nucleos(t)ide treated, HBV-infected subjects in an outpatient hospital setting. HBV DNA levels in untreated people were 3.6 log10 higher than corresponding RNA levels and were stable over 5 years of observation. While only five of 52 treated patients had DNA levels below the lower limit of quantification (10 IU/mL) at the end of follow-up, 13 had HBV RNA levels persistently above this limit, including eight with undetectable DNA. In samples with undetectable core-related antigen we observed a median HBsAg titer 2.7-fold higher than in samples with undetectable RNA (adjusted P = 0.012). Detectable HBV RNA with undetectable HBV DNA was a negative predictor of HBsAg decrease to a level ≤100 IU/mL (P = 0.03). In naive patients the difference between HBV DNA and RNA was higher than previously reported. HBV RNA rapidly decreased during treatment. However, in some cases, it was detectable even after years of effective therapy, being a negative predictor of HBsAg decrease. The investigational RNA assay for use on the Cobas 6800/8800 instruments is a sensitive and standardized method that could be applied in general management of HBV infection. IMPORTANCE This study focused on the quantification of circulating HBV RNA by using a standardized and sensitive assay. Thanks to this system we observed a higher difference between circulating HBV DNA and RNA than previously reported. In treated patients, HBV RNA decreased together with DNA, although some patients presented detectable levels even after years of successful antiviral treatment, suggesting a persistent viral transcription. Of note, the detection of viral RNA when HBV DNA is undetectable was a negative predictor of HBsAg decrease to a level ≤100 IU/mL. This assay could be extremely helpful in HBV patients management to study viral transcription and to identify those treated patients that may achieve sustained viral suppression. 20211108 20220311 2165-0497 Pages: Page: Label: 1 Annotations: Annotation: Subtype: Link Rect: 50, 565, 145, 574 Annotation: Subtype: Link Rect: 417, 144, 513, 151 Annotation: Subtype: Link Rect: 400, 135, 456, 142 Annotation: Subtype: Link Rect: 444, 24, 526, 31 Annotation: Subtype: Link Rect: 508, 683, 544, 719 Page: Label: 2 Annotations: Annotation: Subtype: Link Rect: 87, 595, 92, 604 Destination: 137 Annotation: Subtype: Link Rect: 146, 571, 151, 580 Destination: 137 Annotation: Subtype: Link Rect: 92, 475, 96, 484 Destination: 137 Annotation: Subtype: Link Rect: 166, 463, 170, 472 Destination: 137 Annotation: Subtype: Link Rect: 251, 451, 256, 460 Destination: 137 Annotation: Subtype: Link Rect: 260, 451, 265, 460 Destination: 137 Annotation: Subtype: Link Rect: 363, 415, 368, 424 Destination: 137 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