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Updated: Apr 6, 2026
Use of Viral Entry Assays and Molecular Docking Analysis for the Identification of Antiviral Candidates against Coxsackievirus A16
Published on: July 15, 2019
Development and characterization of a clinical strain of Coxsackievirus A16 and an eGFP infectious clone
Chenglin Deng1, Xiaodan Li, Siqing Liu
1Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, 430071, China.
Insights
Researchers developed a reporter virus from Coxsackievirus A16 (CA16), a cause of hand, foot, and mouth disease (HFMD). This tool aids in studying CA16 infection and evaluating antiviral treatments for HFMD.
Area of Science:
- Virology
- Molecular Biology
- Public Health
Background:
- Coxsackievirus A16 (CA16) is a primary cause of hand, foot, and mouth disease (HFMD), a significant public health concern, especially in Asia.
- Understanding CA16 pathogenesis and replication is crucial for developing effective control strategies.
Purpose of the Study:
- To construct a stable infectious cDNA clone of an epidemic CA16 strain.
- To create a reporter virus (eGFP-CA16) for studying CA16 infection and evaluating antiviral agents.
Main Methods:
- Assembled a stable infectious cDNA clone of an epidemic CA16 strain.
- Engineered a reporter virus (eGFP-CA16) by inserting eGFP into the CA16 genome.
- Generated infectious recombinant viruses by transfecting Vero cells.
Main Results:
- Parental and recombinant viruses exhibited indistinguishable growth characteristics and plaque morphology in vitro.
- eGFP-CA16 showed similar growth trends and EC50 of NITD008 compared to CA16, with a slightly smaller plaque size.
- The stable infectious cDNA clone and reporter virus serve as valuable experimental systems.
Conclusions:
- The developed CA16 infectious clone provides a robust system for studying CA16 pathogenesis and host responses.
- The eGFP-CA16 reporter virus is a powerful tool for monitoring viral infection and assessing antiviral drug efficacy against CA16.
Abstract:
Coxsackievirus A16 (CA16) is one of the major causes of hand, foot, and mouth disease (HFMD) worldwide, which is a common illness that affects children. The frequent occurrence of HFMD outbreaks has become a serious public health problem in Asia. Therefore, it is important to understand the pathogenesis and replication of CA16. In this study, a stable infectious cDNA clone of an epidemic strain of Coxsackievirus A16 (CA16) was assembled, and subsequently a reporter virus (eGFP-CA16) was constructed by inserting the eGFP gene between the 5'-UTR and the N-terminus of VP4, with the addition of a 2A protease cleavage site (ITTLG) at its C-terminus. This was transfected into Vero cells to generate infectious recombinant viruses. The growth characteristics and plaque morphology, in vitro, in mammalian cells were found to be indistinguishable between the parental and recombinant viruses. Although the eGFP-CA16 showed smaller plaque size as compared to recombinant CA16, both were found to exhibit similar growth trends and EC50 of NITD008. In summary, this stable infectious cDNA clone should provide a valuable experimental system to study CA16 infection and host response. The eGFP-CA16 is expected to provide a powerful tool to monitor eGFP expression in infected cells and to evaluate the antiviral activity of potential antiviral agents in the treatment of CA16 infections.
