{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,12]],"date-time":"2026-10-12T20:21:28Z","timestamp":1791836488699,"version":"4.3.4"},"reference-count":68,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2026,1,31]],"date-time":"2026-01-31T00:00:00Z","timestamp":1769817600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Interdisciplinary Construction Program of the College of Laboratory Medicine at Ningxia Medical University","award":["JCXK2025006"],"award-info":[{"award-number":["JCXK2025006"]}]},{"name":"Natural Science Foundation of Ningxia Hui Autonomous Region","award":["2023AAC02057"],"award-info":[{"award-number":["2023AAC02057"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pathogens"],"abstract":"<jats:p>Hepatitis B virus (HBV) pregenomic RNA (pgRNA), transcribed directly from nuclear covalently closed circular DNA (cccDNA), is an essential component in viral replication. The synthesis and encapsidation of pgRNA depend significantly on the transcriptional activity of cccDNA, making serum pgRNA a recently recognized non-invasive biomarker for evaluating cccDNA activity. However, its clinical application is limited by factors including preanalytical variables, methodological inconsistencies in detection, and a lack of standardization in quantification. This review provides an overview of the biological origins of pgRNA and its critical role in the HBV replication cycle, highlighting the stability challenges encountered during the collection, processing, and storage of plasma\/serum samples. Furthermore, it analyzes recent significant advancements in pgRNA detection technologies, encompassing modified reverse transcription quantitative polymerase chain reaction (RT-qPCR), nucleocapsid-captured methodologies, automated testing platforms, multiplex digital PCR, isothermal amplification, and clustered regularly interspaced short palindromic repeats-based assays. A comparison of these technologies revealed that discrepancies in pgRNA quantification arise primarily from variations in sample processing and measurement systems, rather than from inherent biological limitations. Therefore, establishing standardized sample handling procedures, harmonized detection methods, and unified measurement systems is imperative before pgRNA can be reliably applied to monitor treatment, guide cessation decisions, or evaluate cure in chronic hepatitis B.<\/jats:p>","DOI":"10.3390\/pathogens15020153","type":"journal-article","created":{"date-parts":[[2026,2,2]],"date-time":"2026-02-02T09:48:08Z","timestamp":1770025688000},"page":"153","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Challenges and Solutions in pgRNA Measurement: Toward Improved Monitoring of Hepatitis B Therapy"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0009-0004-8776-9687","authenticated-orcid":false,"given":"Zhenkun","family":"Zhu","sequence":"first","affiliation":[{"name":"Department of Clinical Laboratory Medicine, Third Clinical Medical College of Ningxia Medical University, People\u2019s Hospital of Ningxia Hui Autonomous Region, No. 255 Zhengyuan North Street, Jinfeng District, Yinchuan 750002, China"},{"name":"Xiangyang Central Hospital, Affiliated Hospital of Hubei University of Arts and Science, Xiangyang 441021, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jin","family":"Wu","sequence":"additional","affiliation":[{"name":"Department of Clinical Laboratory Medicine, Third Clinical Medical College of Ningxia Medical University, People\u2019s Hospital of Ningxia Hui Autonomous Region, No. 255 Zhengyuan North Street, Jinfeng District, Yinchuan 750002, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jinyuan","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Clinical Laboratory Medicine, Third Clinical Medical College of Ningxia Medical University, People\u2019s Hospital of Ningxia Hui Autonomous Region, No. 255 Zhengyuan North Street, Jinfeng District, Yinchuan 750002, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7784-7854","authenticated-orcid":false,"given":"Tao","family":"Wu","sequence":"additional","affiliation":[{"name":"Department of Clinical Laboratory Medicine, Third Clinical Medical College of Ningxia Medical University, People\u2019s Hospital of Ningxia Hui Autonomous Region, No. 255 Zhengyuan North Street, Jinfeng District, Yinchuan 750002, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,1,31]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"796","DOI":"10.1016\/S2468-1253(22)00124-8","article-title":"Global, regional, and national burden of hepatitis B, 1990\u20132019: A systematic analysis for the Global Burden of Disease Study 2019","volume":"7","author":"Collaborators","year":"2022","journal-title":"Lancet Gastroenterol. Hepatol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1039","DOI":"10.1016\/S0140-6736(22)01468-4","article-title":"Hepatitis B","volume":"401","author":"Jeng","year":"2023","journal-title":"Lancet"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"685","DOI":"10.1016\/j.cld.2007.08.002","article-title":"Molecular virology of hepatitis B virus for clinicians","volume":"11","author":"Block","year":"2007","journal-title":"Clin. Liver Dis."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"185","DOI":"10.1016\/j.antiviral.2018.08.014","article-title":"Host functions used by hepatitis B virus to complete its life cycle: Implications for developing host-targeting agents to treat chronic hepatitis B","volume":"158","author":"Mitra","year":"2018","journal-title":"Antivir. Res."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Hu, J., and Liu, K. (2017). Complete and Incomplete Hepatitis B Virus Particles: Formation, Function, and Application. Viruses, 9.","DOI":"10.3390\/v9030056"},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Shen, S., Xie, Z., Cai, D., Yu, X., Zhang, H., Kim, E.S., Zhou, B., Hou, J., Zhang, X., and Huang, Q. (2020). Biogenesis and molecular characteristics of serum hepatitis B virus RNA. PLoS Pathog., 16.","DOI":"10.1371\/journal.ppat.1008945"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"5937","DOI":"10.1038\/s41598-017-05242-y","article-title":"Baseline value of intrahepatic HBV DNA over cccDNA predicts patient\u2019s response to interferon therapy","volume":"7","author":"Mu","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Allweiss, L., and Dandri, M. (2017). The Role of cccDNA in HBV Maintenance. Viruses, 9.","DOI":"10.3390\/v9060156"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1007\/s10620-017-4842-1","article-title":"Advanced Strategies for Eliminating the cccDNA of HBV","volume":"63","author":"Dong","year":"2018","journal-title":"Dig. Dis. Sci."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1007\/s12072-015-9675-4","article-title":"Asian-Pacific clinical practice guidelines on the management of hepatitis B: A 2015 update","volume":"10","author":"Sarin","year":"2016","journal-title":"Hepatol. Int."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"135","DOI":"10.1016\/j.coviro.2016.06.003","article-title":"HBV cure: Why, how, when?","volume":"18","author":"Levrero","year":"2016","journal-title":"Curr. Opin. Virol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"255","DOI":"10.1111\/jvh.13926","article-title":"Biogenesis of serum HBV RNA and clinical phenomena of serum HBV RNA in chronic hepatitis B patients before and after receiving nucleos(t)ide analogues therapy","volume":"31","author":"Wu","year":"2024","journal-title":"J. Viral Hepat."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"235","DOI":"10.1016\/j.virusres.2007.12.024","article-title":"Hepatitis B viruses: Reverse transcription a different way","volume":"134","author":"Nassal","year":"2008","journal-title":"Virus Res."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"a021386","DOI":"10.1101\/cshperspect.a021386","article-title":"Hepadnavirus Genome Replication and Persistence","volume":"5","author":"Hu","year":"2015","journal-title":"Cold Spring Harb. Perspect. Med."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"3413","DOI":"10.1002\/j.1460-2075.1992.tb05420.x","article-title":"Hepadnaviral assembly is initiated by polymerase binding to the encapsidation signal in the viral RNA genome","volume":"11","author":"Bartenschlager","year":"1992","journal-title":"EMBO J."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"5741","DOI":"10.1128\/jvi.70.9.5741-5750.1996","article-title":"Evidence for activation of the hepatitis B virus polymerase by binding of its RNA template","volume":"70","author":"Tavis","year":"1996","journal-title":"J. Virol."},{"key":"ref_17","first-page":"297","article-title":"Hepatitis B virus morphogenesis","volume":"214","author":"Nassal","year":"1996","journal-title":"Curr. Top. Microbiol. Immunol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"3254","DOI":"10.1128\/jvi.67.6.3254-3263.1993","article-title":"An RNA stem-loop structure directs hepatitis B virus genomic RNA encapsidation","volume":"67","author":"Pollack","year":"1993","journal-title":"J. Virol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"663","DOI":"10.1016\/0092-8674(92)90599-8","article-title":"The reverse transcriptase of hepatitis B virus acts as a protein primer for viral DNA synthesis","volume":"71","author":"Wang","year":"1992","journal-title":"Cell"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"163","DOI":"10.20517\/2394-5079.2016.05","article-title":"Hepatitis B virus molecular biology and pathogenesis","volume":"2","author":"Lamontagne","year":"2016","journal-title":"Hepatoma Res."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"e56","DOI":"10.1038\/emi.2013.56","article-title":"Hepatitis B virus reverse transcriptase: Diverse functions as classical and emerging targets for antiviral intervention","volume":"2","author":"Jones","year":"2013","journal-title":"Emerg. Microbes Infect."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Tu, T., Budzinska, M.A., Shackel, N.A., and Urban, S. (2017). HBV DNA Integration: Molecular Mechanisms and Clinical Implications. Viruses, 9.","DOI":"10.3390\/v9040075"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Schreiner, S., and Nassal, M. (2017). A Role for the Host DNA Damage Response in Hepatitis B Virus cccDNA Formation-and Beyond?. Viruses, 9.","DOI":"10.3390\/v9050125"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1560","DOI":"10.1002\/hep.29800","article-title":"Update on prevention, diagnosis, and treatment of chronic hepatitis B: AASLD 2018 hepatitis B guidance","volume":"67","author":"Terrault","year":"2018","journal-title":"Hepatology"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"66","DOI":"10.1002\/hep.27381","article-title":"Serum hepatitis B virus RNA levels as an early predictor of hepatitis B envelope antigen seroconversion during treatment with polymerase inhibitors","volume":"61","author":"Bartens","year":"2015","journal-title":"Hepatology"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1066","DOI":"10.1093\/infdis\/jiy270","article-title":"Serum HBV RNA as a Predictor of Peginterferon Alfa-2a Response in Patients With HBeAg-Positive Chronic Hepatitis B","volume":"218","author":"Krauel","year":"2018","journal-title":"J. Infect. Dis."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1016\/j.bdq.2018.03.001","article-title":"Improving the standardization of mRNA measurement by RT-qPCR","volume":"15","author":"Sanders","year":"2018","journal-title":"Biomol. Detect. Quantif."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"493","DOI":"10.1128\/JCM.36.2.493-498.1998","article-title":"Stability of human immunodeficiency virus RNA in blood specimens as measured by a commercial PCR-based assay","volume":"36","author":"Sebire","year":"1998","journal-title":"J. Clin. Microbiol."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Gerbehaye, A.-I.d.M.d., Bod\u00e9us, M., Robert, A., Horsmans, Y., and Goubau, P. (2002). Stable hepatitis C virus RNA detection by RT-PCR during four days storage. BMC Infect. Dis., 2.","DOI":"10.1186\/1471-2334-2-22"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1520","DOI":"10.1002\/hep.32479","article-title":"Circulating HBV RNA: From biology to clinical applications","volume":"76","author":"Deng","year":"2022","journal-title":"Hepatology"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"27128","DOI":"10.1038\/s41598-024-77329-2","article-title":"Limited stability of Hepatitis B virus RNA in plasma and serum","volume":"14","author":"Ohlendorf","year":"2024","journal-title":"Sci. Rep."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"4011","DOI":"10.1128\/jvi.63.9.4011-4018.1989","article-title":"Hepatitis B virus transcript produced by RNA splicing","volume":"63","author":"Su","year":"1989","journal-title":"J. Virol."},{"key":"ref_33","first-page":"10","article-title":"In vivo and in vitro expression of defective hepatitis B virus particles generated by spliced hepatitis B virus RNA","volume":"22","author":"Rosmorduc","year":"1995","journal-title":"Hepatology"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"e0099","DOI":"10.1097\/HC9.0000000000000099","article-title":"Measuring hepatitis B pgRNA stability using an updated automated HBV pgRNA assay with increased sensitivity","volume":"7","author":"Anderson","year":"2023","journal-title":"Hepatol. Commun."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"235","DOI":"10.1186\/s12985-021-01706-w","article-title":"Hepatitis B seroconversion revisited: New insights into the natural history of acute hepatitis B virus (HBV) infection from quantitative and highly sensitive assays and novel biomarkers","volume":"18","author":"Kuhns","year":"2021","journal-title":"Virol. J."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"116582","DOI":"10.1016\/j.diagmicrobio.2024.116582","article-title":"A molecular detection method for HBV pgRNA without RNA extraction based on nucleic acid hybridization","volume":"111","author":"Hao","year":"2025","journal-title":"Diagn. Microbiol. Infect. Dis."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"2683","DOI":"10.1007\/s00705-019-04372-0","article-title":"A novel one-step quantitative reverse transcription PCR assay for selective amplification of hepatitis B virus pregenomic RNA from a mixture of HBV DNA and RNA in serum","volume":"164","author":"Gao","year":"2019","journal-title":"Arch. Virol."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Liu, Y., Nie, H., Mao, R., Mitra, B., Cai, D., Yan, R., Guo, J.T., Block, T.M., Mechti, N., and Guo, H. (2017). Interferon-inducible ribonuclease ISG20 inhibits hepatitis B virus replication through directly binding to the epsilon stem-loop structure of viral RNA. PLoS Pathog., 13.","DOI":"10.1371\/journal.ppat.1006296"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"6387","DOI":"10.1128\/JVI.00236-10","article-title":"Inhibition of hepatitis B virus replication by MyD88 involves accelerated degradation of pregenomic RNA and nuclear retention of pre-S\/S RNAs","volume":"84","author":"Li","year":"2010","journal-title":"J. Virol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"86","DOI":"10.1186\/s12985-018-0994-7","article-title":"High serum levels of pregenomic RNA reflect frequently failing reverse transcription in hepatitis B virus particles","volume":"15","author":"Prakash","year":"2018","journal-title":"Virol. J."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"20","DOI":"10.1016\/j.antiviral.2017.07.005","article-title":"NgAgo-gDNA system efficiently suppresses hepatitis B virus replication through accelerating decay of pregenomic RNA","volume":"145","author":"Wu","year":"2017","journal-title":"Antivir. Res."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1016\/j.antiviral.2013.01.006","article-title":"Chronic hepatitis B: What should be the goal for new therapies?","volume":"98","author":"Block","year":"2013","journal-title":"Antivir. Res."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"2106","DOI":"10.1002\/hep.30082","article-title":"Hepatitis B Virus Serum DNA andRNA Levels in Nucleos(t)ide Analog-Treated or Untreated Patients During Chronic and Acute Infection","volume":"68","author":"Butler","year":"2018","journal-title":"Hepatology"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"287","DOI":"10.2144\/000114354","article-title":"Development of a novel hepatitis B virus encapsidation detection assay by viral nucleocapsid-captured quantitative RT-PCR","volume":"59","author":"Ryu","year":"2015","journal-title":"Biotechniques"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"e18557","DOI":"10.1016\/j.heliyon.2023.e18557","article-title":"Evaluation of reverse transcription-polymerase chain reaction and simultaneous amplification and testing for quantitative detection of serum hepatitis B virus RNA","volume":"9","author":"Hu","year":"2023","journal-title":"Heliyon"},{"key":"ref_46","doi-asserted-by":"crossref","unstructured":"Liu, Y., Jiang, M., Xue, J., Yan, H., and Liang, X. (2019). Serum HBV RNA quantification: Useful for monitoring natural history of chronic hepatitis B infection. BMC Gastroenterol., 19.","DOI":"10.1186\/s12876-019-0966-4"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"e27433","DOI":"10.1097\/MD.0000000000027433","article-title":"Analysis of serum hepatitis B virus RNA levels among HBsAg and HBsAb copositive patients and its correlation with HBV DNA","volume":"100","author":"Xiang","year":"2021","journal-title":"Medicine"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"e0214921","DOI":"10.1128\/spectrum.02149-21","article-title":"Standardized Hepatitis B Virus RNA Quantification in Untreated and Treated Chronic Patients: A Promising Marker of Infection Follow-Up","volume":"10","author":"Cortese","year":"2022","journal-title":"Microbiol. Spectr."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"171","DOI":"10.1016\/j.ijid.2020.07.031","article-title":"Efficacy of a combination of HBV RNA and HBeAg in predicting HBeAg seroconversion in patients treated with entecavir for 144 weeks","volume":"99","author":"Wang","year":"2020","journal-title":"Int. J. Infect. Dis."},{"key":"ref_50","doi-asserted-by":"crossref","unstructured":"Pan, J., Tian, Y., Xu, J., Luo, H., Tan, N., Han, Y., Kang, Q., Chen, H., Yang, Y., and Xu, X. (2022). Dynamics of Hepatitis B Virus Pregenomic RNA in Chronic Hepatitis B Patients With Antiviral Therapy Over 9 Years. Front. Med., 9.","DOI":"10.3389\/fmed.2022.851717"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"001856","DOI":"10.1099\/jgv.0.001856","article-title":"An enrichment protocol and analysis pipeline for long read sequencing of the hepatitis B virus transcriptome","volume":"104","author":"Ng","year":"2023","journal-title":"J. Gen. Virol."},{"key":"ref_52","first-page":"001373","article-title":"A PCR assay to quantify patterns of HBV transcription","volume":"102","author":"Magri","year":"2021","journal-title":"J. Gen. Virol."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"4","DOI":"10.1186\/s12985-020-01471-2","article-title":"Serum HBV RNA correlated with intrahepatic cccDNA more strongly than other HBV markers during peg-interferon treatment","volume":"18","author":"Wang","year":"2021","journal-title":"Virol. J."},{"key":"ref_54","doi-asserted-by":"crossref","unstructured":"Sarica, N., Meki, I., Lopez, O., Jay, B., Petitjean, G., and Neuveut, C. (2025). ddOTs: A multiplexed quantitative ddPCR approach for resolving overlapping hepatitis B virus transcripts to decipher cccDNA-driven transcription. bioRxiv, bioRxiv:2025.08.27.672646.","DOI":"10.1101\/2025.08.27.672646"},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"318","DOI":"10.1038\/nature06451","article-title":"Programming biomolecular self-assembly pathways","volume":"451","author":"Yin","year":"2008","journal-title":"Nature"},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"e2303398","DOI":"10.1002\/smll.202303398","article-title":"Digital Recombinase Polymerase Amplification, Digital Loop-Mediated Isothermal Amplification, and Digital CRISPR-Cas Assisted Assay: Current Status, Challenges, and Perspectives","volume":"19","author":"Yin","year":"2023","journal-title":"Small"},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"112075","DOI":"10.1016\/j.microc.2024.112075","article-title":"Detection of HBV pgRNA by catalytic hairpin assembly combined with two Kinds of immunochromatographic strips","volume":"207","author":"Chen","year":"2024","journal-title":"Microchem. J."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"344389","DOI":"10.1016\/j.aca.2025.344389","article-title":"CRISPR\/Cas13a combined with reverse transcription-multienzyme isothermal rapid amplification for hepatitis B virus RNA detection","volume":"1370","author":"Zhu","year":"2025","journal-title":"Anal. Chim. Acta"},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"e0038324","DOI":"10.1128\/jcm.00383-24","article-title":"Rapid and sensitive detection of chikungunya virus using one-tube, reverse transcription, semi-nested multi-enzyme isothermal rapid amplification, and lateral flow dipstick assays","volume":"62","author":"Wu","year":"2024","journal-title":"J. Clin. Microbiol."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"1123","DOI":"10.1038\/s41587-022-01213-5","article-title":"Designing sensitive viral diagnostics with machine learning","volume":"40","author":"Metsky","year":"2022","journal-title":"Nat. Biotechnol."},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"775","DOI":"10.1080\/22221751.2022.2045874","article-title":"A standardized assay for the quantitative detection of serum HBV RNA in chronic hepatitis B patients","volume":"11","author":"Yu","year":"2022","journal-title":"Emerg. Microbes Infect."},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"114640","DOI":"10.1016\/j.jviromet.2022.114640","article-title":"Development of a sensitive, multi-assay platform to monitor low levels of HBV DNA and pgRNA in patients with chronic hepatitis B virus infection","volume":"311","author":"Yan","year":"2023","journal-title":"J. Virol. Methods"},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"847","DOI":"10.1016\/j.jhep.2017.05.008","article-title":"Hepatitis B cure: From discovery to regulatory approval","volume":"67","author":"Lok","year":"2017","journal-title":"J. Hepatol."},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1016\/j.jviromet.2004.05.012","article-title":"Development of a one tube-one step RT-PCR protocol for the detection of seven viroids in four genera: Apscaviroid, Hostuviroid, Pelamoviroid and Pospiviroid","volume":"121","author":"Ragozzino","year":"2004","journal-title":"J. Virol. Methods"},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"e2051","DOI":"10.1002\/rmv.2051","article-title":"Pregenomic RNA: How to assist the management of chronic hepatitis B?","volume":"29","author":"Wu","year":"2019","journal-title":"Rev. Med. Virol."},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"22","DOI":"10.1186\/s12985-022-01749-7","article-title":"Serum hepatitis B virus RNA detectability, composition and clinical significance in patients with ab initio hepatitis B e antigen negative chronic hepatitis B","volume":"19","author":"Laras","year":"2022","journal-title":"Virol. J."},{"key":"ref_67","doi-asserted-by":"crossref","first-page":"3688","DOI":"10.1002\/jmv.26529","article-title":"Distribution and factors associated with serum HBV pregenomic RNA levels in Chinese chronic hepatitis B patients","volume":"93","author":"Wang","year":"2021","journal-title":"J. Med. Virol."},{"key":"ref_68","doi-asserted-by":"crossref","first-page":"189","DOI":"10.1002\/hep4.1633","article-title":"Persistent Control of Hepatitis B Virus and Hepatitis Delta Virus Infection Following REP 2139-Ca and Pegylated Interferon Therapy in Chronic Hepatitis B Virus\/Hepatitis Delta Virus Coinfection","volume":"5","author":"Bazinet","year":"2020","journal-title":"Hepatol. Commun."}],"container-title":["Pathogens"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-0817\/15\/2\/153\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,2,2]],"date-time":"2026-02-02T10:01:37Z","timestamp":1770026497000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-0817\/15\/2\/153"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,1,31]]},"references-count":68,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2026,2]]}},"alternative-id":["pathogens15020153"],"URL":"https:\/\/doi.org\/10.3390\/pathogens15020153","relation":{},"ISSN":["2076-0817"],"issn-type":[{"value":"2076-0817","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,1,31]]}}}