Construction and characterization of an infectious clone of coxsackievirus A6 that showed high virulence in neonatal mice.
Atypical hand, foot, and mouth disease (aHFMD) outbreaks have been frequently reported worldwide in recent years. It is believed that coxsackievirus A6 (CA6) is the major pathogen for aHFMD. Studies regarding CA6 infection are limited and the genetic mechanism for the high pathogenicity of some new CA6 variants is still unclear. Infectious clones are powerful tools for studying the genetic mechanisms of RNA viruses. In this study, we describe the construction of a full-length cDNA clone of CA6 strain TW-2007-00141. The whole genome of CA6 was amplified in a single step and ligated into a plasmid vector through an efficient cloning method, Gibson assembly. The whole genome sequence of CA6 strain TW-2007-00141 was determined and phylogenetic analysis indicated that it shared a high degree of similarity (≥94%) with the CA6 strains found in Taiwan in 2009. The infectious clone of CA6 viruses were recovered by transfection into 293FT cells and showed similar biological properties to the parental virus. Viral particles were purified by CsCl isopycnic centrifugation, and two types of viral particles were observed under transmission electron microscopy. The rescued virus showed high virulence in one-day-old suckling mice. This clone may be useful for establishing animal models for the evaluation of CA6 vaccine efficiency in future.
State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Xiamen University, Xiamen 361102, China.
Lisheng
L
Yang
State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Xiamen University, Xiamen 361102, China.
Shuxuan
S
Li
State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Xiamen University, Xiamen 361102, China; National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health, Xiamen University, Xiamen 361102, China.
Yajing
Y
Liu
State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Xiamen University, Xiamen 361102, China.
Wangheng
W
Hou
National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health, Xiamen University, Xiamen 361102, China.
Qiaona
Q
Lin
National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health, Xiamen University, Xiamen 361102, China.
Huan
H
Zhao
National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health, Xiamen University, Xiamen 361102, China.
Longfa
L
Xu
State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Xiamen University, Xiamen 361102, China.
Delei
D
He
Beijing Wantai Biological Pharmacy Enterprise, Beijing 102206, China.
Xiangzhong
X
Ye
Department of Microbiology and Molecular Genetics, New Jersey Medical School, Rutgers University, 225 Warren Street, Newark, NJ 070101, USA.
Hua
H
Zhu
State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Xiamen University, Xiamen 361102, China; National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health, Xiamen University, Xiamen 361102, China. Electronic address: tcheng@xmu.edu.cn.
Tong
T
Cheng
State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Xiamen University, Xiamen 361102, China; National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health, Xiamen University, Xiamen 361102, China.
Ningshao
N
Xia
D000818:Animals; D000831:Animals, Newborn; D003001:Cloning, Molecular; D016000:Cluster Analysis; D003384:Coxsackievirus Infections / Q000473:pathology* / Q000821:virology*; D004195:Disease Models, Animal; D004770:Enterovirus / Q000235:genetics / Q000302:isolation & purification / Q000472:pathogenicity* / Q000648:ultrastructure; D016679:Genome, Viral; D006801:Humans; D008807:Mice, Inbred BALB C; D046529:Microscopy, Electron, Transmission; D008969:Molecular Sequence Data; D021141:Nucleic Acid Amplification Techniques; D010802:Phylogeny; D010957:Plasmids; D012367:RNA, Viral / Q000235:genetics; D059386:Reverse Genetics; D017422:Sequence Analysis, DNA; D017385:Sequence Homology; D013624:Taiwan; D014771:Virion / Q000648:ultrastructure; D014774:Virulence
D012367:RNA, Viral
Animal model; Coxsackievirus A6; Enterovirus; HFMD; Infectious clone