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Sci Rep. 2017 Jan 12;7:40199. doi: 10.1038/srep40199.

A novel toolbox for the in vitro assay of hepatitis D virus infection.

Author information

1
State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Science &School of Public Health, Xiamen University, Xiamen 361102, PR China.
2
National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Life Science &School of Public Health, Xiamen University, Xiamen 361102, PR China.
3
National Taiwan University College of Medicine, National Taiwan University, Taipei 10051, Taiwan.

Abstract

Hepatitis D virus (HDV) is a defective RNA virus that requires the presence of hepatitis B virus (HBV) for its life cycle. The in vitro HDV infection system is widely used as a surrogate model to study cellular infection with both viruses owing to its practical feasibility. However, previous methods for running this system were less efficient for high-throughput screening and large-scale studies. Here, we developed a novel method for the production of infectious HDV by adenoviral vector (AdV)-mediated transduction. We demonstrated that the AdV-based method yields 10-fold higher viral titers than the transient-transfection approach. The HDV-containing supernatant derived from AdV-infected Huh7 cells can be used as the inoculum in infectivity assays without requiring further concentration prior to use. Furthermore, we devloped a chemiluminescent immunoassay (HDV-CLEIA) to quantitatively determine intracellular HDAg with a dynamic range of 5-11,000 pg/mL. HDV-CLEIA can be used as an alternative approach to assess HDV infection. The advantages of our updated methodology were demonstrated through in vitro HDV infection of HepaRG cells and by evaluating the neutralization activity using antibodies that target various regions of the HBV/HDV envelope proteins. Together, the methods presented here comprise a novel toolbox of in vitro assays for studying HDV infection.

PMID:
28079152
PMCID:
PMC5228157
DOI:
10.1038/srep40199
[Indexed for MEDLINE]
Free PMC Article

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