Autor: |
Takkenberg, R. B., Menting, S., Beld, M. G. H. M. |
Přispěvatelé: |
Amsterdam Gastroenterology Endocrinology Metabolism, Gastroenterology and Hepatology, Dermatology |
Jazyk: |
angličtina |
Rok vydání: |
2012 |
Předmět: |
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Zdroj: |
Methods in molecular biology (Clifton, N.J.), 903, 113-128. Humana Press |
ISSN: |
1064-3745 |
Popis: |
Hepatitis B virus (HBV) covalently closed circular DNA (cccDNA) serves as a template for viral replication and plays a role in persistence of HBV infection. The origin and significance of cccDNA in plasma, however, are not well understood. A sensitive, specific, and reproducible real-time PCR for detection and quantitation of cccDNA in plasma of chronic hepatitis B patients was developed and validated. Four HBV DNA reference panels and 96 plasma samples of chronic hepatitis B patients are analyzed. Results are compared with total HBV DNA levels. This cccDNA assay had a lower limit of detection at 15 copies/PCR, a lower limit of quantitation at 91 copies/PCR, and a correlation coefficient (R) of 0.98 (p < 0.0001). HBV cccDNA can be detected in two of four international panels. Significant correlation is found between cccDNA and total HBV DNA levels in both panels (R = 0.96 and R = 0.43) and in samples of the chronic hepatitis B patients (R = 0.88, p < 0.0001). In 57 % of these samples cccDNA can be detected. Mean level of cccDNA is 0.16 % of total HBV load. Plasma HBV cccDNA levels are higher in HBeAg-positive samples than in HBeAg-negative samples (p < 0.0001). Total HBV DNA levels and HBV genotype do not influence cccDNA detection |
Databáze: |
OpenAIRE |
Externí odkaz: |
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