Validation of a sensitive and specific real-time PCR for detection and quantitation of hepatitis B virus covalently closed circular DNA in plasma of chronic hepatitis B patients

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, is not well understood. A sensitive, specific, and reproducible real‐time PCR for dete...

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Bibliographic Details
Published in:Journal of medical virology 2009-06, Vol.81 (6), p.988-995
Main Authors: Takkenberg, R.B., Zaaijer, H.L., Molenkamp, R., Menting, S., Terpstra, V., Weegink, C.J., Dijkgraaf, M.G.W., Jansen, P.L.M., Reesink, H.W., Beld, M.G.H.M.
Format: Article
Language:English
Subjects:
Adult
Biological and medical sciences
cccDNA
DNA, Circular - blood
DNA, Circular - genetics
DNA, Viral - blood
DNA, Viral - genetics
Fundamental and applied biological sciences. Psychology
HBV
Hepatitis B Surface Antigens - blood
Hepatitis B virus
Hepatitis B virus - genetics
Hepatitis B, Chronic - virology
Human viral diseases
Humans
Infectious diseases
Medical sciences
Microbiology
Middle Aged
Miscellaneous
plasma
Plasma - virology
Polymerase Chain Reaction - methods
quantitation
real-time PCR
Sensitivity and Specificity
Viral diseases
Viral Load
Virology
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Summary: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, is 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 were analyzed. Results were compared with total HBV DNA levels, individual ALT levels and the Histology Activity Index (HAI). 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 
ISSN:0146-6615
1096-9071
DOI:10.1002/jmv.21477