Abundance of Noncircular Intrahepatic Hepatitis B Virus DNA May Reflect Frequent Integration Into Human DNA in Chronically Infected Patients
| Autor: | Helene Norder, Gustaf E. Rydell, Gunnar Norkrans, Simon B. Larsson, Kasthuri Prakash, Magnus Lindh, Kristoffer Hellstrand, Maria Andersson |
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| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Hepatitis B virus HBsAg Biology medicine.disease_cause Genome 03 medical and health sciences chemistry.chemical_compound Hepatitis B Chronic 0302 clinical medicine Antigen medicine Humans Immunology and Allergy Digital polymerase chain reaction Hepatitis B e Antigens Nuclease Hepatitis B Surface Antigens medicine.diagnostic_test virus diseases Hepatitis B Virology digestive system diseases 030104 developmental biology Infectious Diseases Liver chemistry Liver biopsy DNA Viral biology.protein 030211 gastroenterology & hepatology DNA Circular DNA |
| Zdroj: | The Journal of Infectious Diseases. 225:1982-1990 |
| ISSN: | 1537-6613 0022-1899 |
| DOI: | 10.1093/infdis/jiaa572 |
| Popis: | Background Hepatitis B virus (HBV) integration has implications for cancer development and surface antigen (HBsAg) production, but methods to quantify integrations are lacking. The aim of this study was to develop a droplet digital PCR (ddPCR) assay discriminating between circular and integrated HBV DNA, and to relate the distribution between the two forms to other HBV markers. Methods ddPCR with primers spanning the typical linearization breakpoint in the HBV genome allowed for quantification of the absolute copy numbers of total and circular HBV DNA, and calculation of linear HBV DNA. Results Analysis of 70 liver biopsies from patients with chronic HBV infection revealed that the fraction of linear HBV DNA, which includes integrations, was higher in HBeAg-negative patients than HBeAg-positive. The ratio between HBsAg and HBV DNA levels in serum correlated with the intrahepatic proportion of linear HBV DNA. Furthermore, ddPCR experiments on serum samples and experiments with nuclease indicated the contribution of encapsidated double-stranded linear DNA and replication intermediates to be limited. Conclusions The degree of integration of intrahepatic HBV DNA in the HBeAg-negative stage may be higher than previously anticipated, and integrated DNA may explain the persistence of high HBsAg serum levels in patients with low HBV DNA levels. |
| Databáze: | OpenAIRE |
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