Hepatitis B Virus RNA Profiles in Liver Biopsies by Digital Polymerase Chain Reaction
Replication of hepatitis B virus (HBV) originates from covalently closed circular DNA (cccDNA) and involves reverse transcription of pregenomic RNA (pgRNA), which is also called core RNA and encodes the capsid protein. The RNA coding for hepatitis B surface antigen (HBsAg) in the envelope of viral o...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Wolters Kluwer Health/LWW
2020-07-01
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| Series: | Hepatology Communications |
| Online Access: | https://doi.org/10.1002/hep4.1507 |
| _version_ | 1811174060961300480 |
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| author | Kasthuri Prakash Simon B. Larsson Gustaf E. Rydell Maria E. Andersson Johan Ringlander Gunnar Norkrans Heléne Norder Magnus Lindh |
| author_facet | Kasthuri Prakash Simon B. Larsson Gustaf E. Rydell Maria E. Andersson Johan Ringlander Gunnar Norkrans Heléne Norder Magnus Lindh |
| author_sort | Kasthuri Prakash |
| collection | DOAJ |
| description | Replication of hepatitis B virus (HBV) originates from covalently closed circular DNA (cccDNA) and involves reverse transcription of pregenomic RNA (pgRNA), which is also called core RNA and encodes the capsid protein. The RNA coding for hepatitis B surface antigen (HBsAg) in the envelope of viral or subviral particles is produced from cccDNA or from HBV DNA integrated into the host genome. Because only cccDNA can generate the core and the 3′ redundancy regions of HBV RNA, we aimed to clarify to what extent such HBV integrations are expressed by quantifying the different HBV RNA species in liver tissue. Digital droplet polymerase chain reaction (ddPCR) was employed to quantify six HBV RNA targets in 76 liver biopsies from patients with chronic infection, comprising 14 who were hepatitis B e antigen (HBeAg) positive and 62 who were HBeAg negative. In patients who were HBeAg negative, HBV RNA from the S RNA region was >1.6 log10 units higher than in the core and 3′ redundancy regions (P < 0.0001), indicating that >90% of S RNA was integration derived. HBeAg‐negative samples showed 10 times lower levels of pgRNA (5′ core) compared with core RNA (3′ part of core; P < 0.0001), suggesting that a large proportion of core RNA might have a downstream shift of the transcription starting point. In multiple regression analysis, HBV DNA levels in serum were most strongly dependent on pgRNA. Conclusion: In patients who were HBeAg negative, integration‐derived S RNA seemed to predominate and a large proportion of the core RNA lacked the 5′ part. Because this part comprises the down‐regulator of transcription 1 sequences, which are necessary for virus production (plus strand translocation), the finding might help to explain the low level of HBV DNA in serum that frequently is observed in patients with chronic HBV infection who are HBeAg negative. |
| first_indexed | 2024-04-10T17:57:02Z |
| format | Article |
| id | doaj.art-29e8428a4dc140b99b48ab94b24dd0d3 |
| institution | Directory Open Access Journal |
| issn | 2471-254X |
| language | English |
| last_indexed | 2024-04-10T17:57:02Z |
| publishDate | 2020-07-01 |
| publisher | Wolters Kluwer Health/LWW |
| record_format | Article |
| series | Hepatology Communications |
| spelling | doaj.art-29e8428a4dc140b99b48ab94b24dd0d32023-02-02T17:27:44ZengWolters Kluwer Health/LWWHepatology Communications2471-254X2020-07-014797398210.1002/hep4.1507Hepatitis B Virus RNA Profiles in Liver Biopsies by Digital Polymerase Chain ReactionKasthuri Prakash0Simon B. Larsson1Gustaf E. Rydell2Maria E. Andersson3Johan Ringlander4Gunnar Norkrans5Heléne Norder6Magnus Lindh7Department of Infectious Diseases Sahlgrenska Academy University of Gothenburg Gothenburg SwedenDepartment of Infectious Diseases Sahlgrenska Academy University of Gothenburg Gothenburg SwedenDepartment of Infectious Diseases Sahlgrenska Academy University of Gothenburg Gothenburg SwedenDepartment of Infectious Diseases Sahlgrenska Academy University of Gothenburg Gothenburg SwedenDepartment of Infectious Diseases Sahlgrenska Academy University of Gothenburg Gothenburg SwedenDepartment of Infectious Diseases Sahlgrenska Academy University of Gothenburg Gothenburg SwedenDepartment of Infectious Diseases Sahlgrenska Academy University of Gothenburg Gothenburg SwedenDepartment of Infectious Diseases Sahlgrenska Academy University of Gothenburg Gothenburg SwedenReplication of hepatitis B virus (HBV) originates from covalently closed circular DNA (cccDNA) and involves reverse transcription of pregenomic RNA (pgRNA), which is also called core RNA and encodes the capsid protein. The RNA coding for hepatitis B surface antigen (HBsAg) in the envelope of viral or subviral particles is produced from cccDNA or from HBV DNA integrated into the host genome. Because only cccDNA can generate the core and the 3′ redundancy regions of HBV RNA, we aimed to clarify to what extent such HBV integrations are expressed by quantifying the different HBV RNA species in liver tissue. Digital droplet polymerase chain reaction (ddPCR) was employed to quantify six HBV RNA targets in 76 liver biopsies from patients with chronic infection, comprising 14 who were hepatitis B e antigen (HBeAg) positive and 62 who were HBeAg negative. In patients who were HBeAg negative, HBV RNA from the S RNA region was >1.6 log10 units higher than in the core and 3′ redundancy regions (P < 0.0001), indicating that >90% of S RNA was integration derived. HBeAg‐negative samples showed 10 times lower levels of pgRNA (5′ core) compared with core RNA (3′ part of core; P < 0.0001), suggesting that a large proportion of core RNA might have a downstream shift of the transcription starting point. In multiple regression analysis, HBV DNA levels in serum were most strongly dependent on pgRNA. Conclusion: In patients who were HBeAg negative, integration‐derived S RNA seemed to predominate and a large proportion of the core RNA lacked the 5′ part. Because this part comprises the down‐regulator of transcription 1 sequences, which are necessary for virus production (plus strand translocation), the finding might help to explain the low level of HBV DNA in serum that frequently is observed in patients with chronic HBV infection who are HBeAg negative.https://doi.org/10.1002/hep4.1507 |
| spellingShingle | Kasthuri Prakash Simon B. Larsson Gustaf E. Rydell Maria E. Andersson Johan Ringlander Gunnar Norkrans Heléne Norder Magnus Lindh Hepatitis B Virus RNA Profiles in Liver Biopsies by Digital Polymerase Chain Reaction Hepatology Communications |
| title | Hepatitis B Virus RNA Profiles in Liver Biopsies by Digital Polymerase Chain Reaction |
| title_full | Hepatitis B Virus RNA Profiles in Liver Biopsies by Digital Polymerase Chain Reaction |
| title_fullStr | Hepatitis B Virus RNA Profiles in Liver Biopsies by Digital Polymerase Chain Reaction |
| title_full_unstemmed | Hepatitis B Virus RNA Profiles in Liver Biopsies by Digital Polymerase Chain Reaction |
| title_short | Hepatitis B Virus RNA Profiles in Liver Biopsies by Digital Polymerase Chain Reaction |
| title_sort | hepatitis b virus rna profiles in liver biopsies by digital polymerase chain reaction |
| url | https://doi.org/10.1002/hep4.1507 |
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