{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,23]],"date-time":"2026-03-23T14:52:56Z","timestamp":1774277576553,"version":"3.50.1"},"reference-count":38,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2026,2,13]],"date-time":"2026-02-13T00:00:00Z","timestamp":1770940800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,2,13]],"date-time":"2026-02-13T00:00:00Z","timestamp":1770940800000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100002794","name":"Cancerfonden","doi-asserted-by":"publisher","award":["CAN 20 1280 PjF"],"award-info":[{"award-number":["CAN 20 1280 PjF"]}],"id":[{"id":"10.13039\/501100002794","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Liver International"],"published-print":{"date-parts":[[2026,3]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>Hepatitis B virus (HBV) genomes integrated into human DNA significantly contribute to surface antigen (HBsAg) production and may drive hepatocellular carcinoma (HCC). Long\u2010read sequencing methods like Nanopore offer advantages over short\u2010read next\u2010generation sequencing (NGS) by providing continuous reads of whole transcripts, but their application to HBV integration analysis remains limited.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Objective<\/jats:title>\n                    <jats:p>To develop and apply a method combining semi\u2010nested PCR with Nanopore sequencing to analyse HBV transcripts, including canonical RNA, HBV\u2013human fusion transcripts, and spliced forms in patients with HBV\u2010 or hepatitis D virus (HDV)\u2010induced liver disease.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>Nine liver\u2010transplanted patients with HBV\u2010 or HDV\u2010related cirrhosis or HCC were studied. Semi\u2010nested PCR was used to amplify all HBV transcripts, followed by Nanopore sequencing. The approach allowed differentiation between canonical (cccDNA\u2010derived) and fusion transcripts. Reads containing the 3\u2032 redundancy beyond nucleotide 1826, exclusive to cccDNA\u2010derived RNA, were quantified to determine the source of HBV RNA.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>Unique and total HBV\u2010human fusion RNA reads correlated with serum levels of HBV DNA and HBsAg. Integration\u2010derived RNA accounted for a median of 97% (range: 16%\u2013100%) of HBV RNA. PreS1 RNA levels were much lower than preS2 but sufficient for HDV particle production in an HDV patient without cccDNA\u2010derived transcripts.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>This method enables a simplified and comprehensive analysis of HBV transcripts. The results highlight the predominance of integration\u2010derived RNA and support the presence of cccDNA\u2010independent hepatitis D virus production. Nanopore sequencing offers valuable insights into HBV and HDV biology, supporting its role in understanding viral pathogenesis and therapeutic targeting.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1111\/liv.70521","type":"journal-article","created":{"date-parts":[[2026,2,13]],"date-time":"2026-02-13T08:49:52Z","timestamp":1770972592000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Novel Long\u2010Read Sequencing Method for Characterisation of Hepatitis B Transcripts Show High Expression of Chimeric\n                    <scp>HBV<\/scp>\n                    \/Human\n                    <scp>RNA<\/scp>"],"prefix":"10.1111","volume":"46","author":[{"given":"Joakim Bedner","family":"Stenb\u00e4ck","sequence":"first","affiliation":[{"name":"Department of Infectious Diseases Institute of Biomedicine, Sahlgrenska Academy, University of Gothenburg  Gothenburg Sweden"},{"name":"Department of Clinical Microbiology Sahlgrenska University Hospital  Gothenburg Sweden"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8483-6504","authenticated-orcid":false,"given":"Johan","family":"Ringlander","sequence":"additional","affiliation":[{"name":"Department of Infectious Diseases Institute of Biomedicine, Sahlgrenska Academy, University of Gothenburg  Gothenburg Sweden"},{"name":"Department of Clinical Microbiology Sahlgrenska University Hospital  Gothenburg Sweden"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0005-7114","authenticated-orcid":false,"given":"Maria","family":"Andersson","sequence":"additional","affiliation":[{"name":"Department of Infectious Diseases Institute of Biomedicine, Sahlgrenska Academy, University of Gothenburg  Gothenburg Sweden"},{"name":"Department of Clinical Microbiology Sahlgrenska University Hospital  Gothenburg 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