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Since HBV RNAs are expressed from cccDNA or HBV integrated sequences, independently of viral genome replication, levels of HBV RNAs in plasma may remain high following treatment with nucleos(t)ide analogue. Thus, HBV RNAs have been proposed to be used as a viral biomarker for treatment outcome and disease progression. Recent investigations of plasma HBV RNAs described the presence of full length as well as subgenomic forms of RNA. To support the usage of plasma HBV RNAs as a viral biomarker, further understanding of HBV RNA composition in clinical samples is needed. Here, sequence of extracellular HBV RNAs was characterized in plasma samples of patients with chronic HBV infection using two independent RNA amplification methods that do not use HBV\u2010specific primers for amplification: total RNA (NuGEN RNAseq) and mRNA (TruSeq RNAseq). Sequencing coverage was obtained across the full length of HBV genome for both methods, confirming the presence of full\u2010length HBV RNA in plasma. The sequence of HBV RNA was nearly identical to plasma HBV DNA sequence in each sample with only 0\u201314 (median 4) mismatches over 3\u00a0kb. Thus, sequence of HBV RNA plasma reflects the intrahepatic viral reservoir and can be used for monitoring of sequence variants such as resistance in clinical trials. Additionally, RNA splice forms, different polyA tails start positions and presence of HBV\u2010human chimeric transcript were identified.<\/jats:p>","DOI":"10.1111\/jvh.13760","type":"journal-article","created":{"date-parts":[[2022,10,8]],"date-time":"2022-10-08T09:45:06Z","timestamp":1665222306000},"page":"29-38","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Sequence characterization of extracellular <scp>HBV RNA<\/scp> in patient plasma"],"prefix":"10.1111","volume":"30","author":[{"given":"Silvia","family":"Chang","sequence":"first","affiliation":[{"name":"Gilead Sciences, Inc.  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