Diagnosis and Treatment of Viral Diseases
Recombinant Crimean-Congo hemorrhagic fever virus with Nanoluciferase reporter and its potential application for rapid antiviral compounds screening
Han Xia, Zhiming Yuan, Dennis Bente
Published 2018-12-30
Cite as Chinese J Exp Clin Virol, 2018, 32(6): 620-624. DOI: 10.3760/cma.j.issn.1003-9279.2018.06.013
Abstract
ObjectiveTo construct the recombinant Crimean-Congo hemorrhagic fever virus (CCHFV) which can express the secreted nanoluciferase (NanoLuc) and investigate its potential application for rapid antiviral compounds screening.
MethodsThe ORF of NanoLuc and the mucin encoded by the M segment of CCHFV were merged, and the recombinant CCHFV (rCCHFV) was rescued through reverse genetic system. Then rCCHFV was used to evaluate the antiviral effect for ribavirin and Furin inhibitor in vitro.
ResultsThe rCCHFV_mucin_NLuc with NanoLuc reporter was obtained, and the relative light unit (RLU) which can reflect NanoLuc activity was positively correlated with median tissue culture infective dose (TCID50) in the infected cell supernatant (cor=0.998, P=0.001). When the concentration for the compounds was 10 μmol/L, there was no significant difference for the NanoLuc activity in the infected cell supernatant between Furin inhibitor and ribavirin (P > 0.1) from day 1 to 3 after treatment. But at day 4, the NanoLuc activity in Fruin inhibitor treated group was significantly higher than that of ribavirin treated group (P=0.001), and no significant difference was found between the Furin inhibitor and untreated group (P > 0.1).
ConclusionsThe rCCHFV with NanoLuc reporter was recovered successfully and it could be used for the primary rapid screening of antiviral compounds in future.
Key words:
Luciferase; Crimean-Congo hemorrhagic fever virus; Antiviral compound; Screening
Contributor Information
Han Xia
Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China
Zhiming Yuan
Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China
Dennis Bente
Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston 77555, USA