Construction and characterization of an infectious clone of coxsackievirus A16

• Main Authors: Liu Fei, Liu Qingwei, Cai Yicun, Leng Qibin, Huang Zhong
• Format: Article
• Language: English
• Published: BMC 2011-12-01
• Series: Virology Journal
• Subjects: Coxsackievirus A16; Infectious cDNA clone; <it>In vitro </it>transcription; Recovered virus
• Online Access: http://www.virologyj.com/content/8/1/534

<p>Abstract</p> <p>Background</p> <p>Coxsackievirus A16 (CVA16) is a member of the <it>Enterovirus </it>genus of the <it>Picornaviridae </it>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.</p> <p>Results</p> <p>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 <it>via </it>a native <it>Xba</it>I 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 <it>in vitro </it>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.</p> <p>Conclusions</p> <p>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.</p>