Construction of cDNA infectious clones of EV71 highly-pathogenic and cell-culture-adapted strains.
- Author:
Yong-xin ZHANG
;
Xiao-yu LI
;
Yu-ming HUANG
;
Yong-dong ZHOU
;
Sheng-li BI
;
Shan CEN
- Publication Type:Journal Article
- MeSH:
Animals;
Cercopithecus aethiops;
DNA, Complementary;
Enterovirus A, Human;
genetics;
growth & development;
isolation & purification;
pathogenicity;
Hand, Foot and Mouth Disease;
virology;
Humans;
Phylogeny;
Vero Cells;
Virus Cultivation
- From:
Chinese Journal of Virology
2014;30(6):605-613
- CountryChina
- Language:Chinese
-
Abstract:
The highly-pathogenic EV71 strain is the primary cause of mortality in hand-foot-and-mouth disease (HFMD) associated with neurological symptoms, for which no clinically effective drugs or vaccines exist. This study aimed to construct infectious cDNA clones of the EV71 highly-pathogenic strain and the cell-culture adapted strain using a reverse genetics approach. The genomic RNAs of EV71 parent strains were used as the templates for RT-PCR amplification, and then the PCR products that overlapped the full-length genome were connected into the pBR322 vector to produce infectious clones of pEV71 (HP) and pEV71 (CCA), respectively. The results showed that the HP strain propagated much more quickly than the CCA strain. The rescued viruses derived from the infectious clones not only maintained their consistency with their parent strains in terms of genomic sequences, but also retained their respective biological phenotypes. This research will contribute to our understanding of EV71 pathogenesis and the development of novel vaccines against HFMD.
