Recombinant Crimean-Congo hemorrhagic fever virus with Nanoluciferase reporter and its potential application for rapid antiviral compounds screening
10.3760/cma.j.issn.1003-9279.2018.06.013
- VernacularTitle: NanoLuc标记的重组克里米亚－刚果出血热病毒及其初步用于抗病毒药物筛选
- Author:
Han XIA
1
;
Zhiming YUAN
1
;
Dennis BENTE
2
Author Information
1. Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China
2. Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston 77555, USA
- Publication Type:Journal Article
- Keywords:
Luciferase;
Crimean-Congo hemorrhagic fever virus;
Antiviral compound;
Screening
- From:
Chinese Journal of Experimental and Clinical Virology
2018;32(6):620-624
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To 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.
Methods:The 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.
Results:The 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).
Conclusions:The rCCHFV with NanoLuc reporter was recovered successfully and it could be used for the primary rapid screening of antiviral compounds in future.
