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Updated: May 26, 2026
Use of Viral Entry Assays and Molecular Docking Analysis for the Identification of Antiviral Candidates against Coxsackievirus A16
Published on: July 15, 2019
Construction and characterization of an infectious clone of coxsackievirus A16
Fei Liu1, Qingwei Liu, Yicun Cai
1Key Laboratory of Molecular Virology & Immunology, Institute Pasteur of Shanghai, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200025, China.
Insights
Researchers created the first infectious Coxsackievirus A16 (CVA16) cDNA clone. This breakthrough enables enhanced studies on CVA16 virology and the development of new vaccines for hand, foot, and mouth disease.
Area of Science:
- Virology
- Molecular Biology
- Infectious Diseases
Background:
- Coxsackievirus A16 (CVA16) is a primary cause of hand, foot, and mouth disease (HFMD) in children.
- Limited knowledge exists regarding CVA16 replication, structure, and virulence factors.
- Difficulties in manipulating the CVA16 RNA genome hinder research.
Purpose of the Study:
- To construct and characterize the first infectious cDNA clone of CVA16.
- To facilitate future virological investigations of CVA16.
- To support the development of CVA16 vaccines.
Main Methods:
- Amplification of two overlapping cDNA fragments from CVA16 RNA using RT-PCR.
- Ligation of cDNA fragments to create a full-length CVA16 cDNA clone with T7 promoter and poly(A) tail.
- Transfection of RD cells with transcribed RNA to recover infectious CVA16.
Main Results:
- A full-length infectious cDNA clone of CVA16 was successfully constructed.
- Transfection with transcribed RNA led to the recovery of infectious CVA16 in cell culture.
- The recovered CVA16 was functionally and genetically identical to the parent strain.
Conclusions:
- The first infectious cDNA clone of CVA16 has been successfully created and characterized.
- This infectious clone is a valuable tool for advancing CVA16 research.
- The availability of this clone will accelerate CVA16 vaccine development.
Background:
Coxsackievirus A16 (CVA16) is a member of the Enterovirus genus of the Picornaviridae family and it is a major etiological agent of hand, foot, and mouth disease (HFMD), which is a common illness affecting children. CVA16 possesses a single-stranded positive-sense RNA genome containing approximately 7410 bases. Current understanding of the replication, structure and virulence determinants of CVA16 is very limited, partly due to difficulties in directly manipulating its RNA genome.
Results:
Two overlapping cDNA fragments were amplified by RT-PCR from the genome of the shzh05-1 strain of CVA16, encompassing the nucleotide regions 1-4392 and 4381-7410, respectively. These two fragments were then joined via a native XbaI site to yield a full-length cDNA. A T7 promoter and poly(A) tail were added to the 5' and 3' ends, respectively, forming a full CVA16 cDNA clone. Transfection of RD cells in vitro with RNA transcribed directly from the cDNA clone allowed the recovery of infectious virus in culture. The CVA16 virus recovered from these cultures was functionally and genetically identical to its parent strain.
Conclusions:
We report the first construction and characterization of an infectious cDNA clone of CVA16. The availability of this infectious clone will greatly enhance future virological investigations and vaccine development for CVA16.
