Zhang Yong-xin, Li Xiao-yu, Huang Yu-ming, Zhou Yong-dong, Bi Sheng-li, Cen Shan
Bing Du Xue Bao. 2014 Nov;30(6):605-13.
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.
高致病性肠道病毒71型(EV71)毒株是伴有神经症状的手足口病(HFMD)死亡的主要原因,目前尚无临床有效的药物或疫苗。本研究旨在采用反向遗传学方法构建EV71高致病性毒株和细胞培养适应株的感染性cDNA克隆。以EV71亲本毒株的基因组RNA为模板进行RT-PCR扩增,然后将与全长基因组重叠的PCR产物连接到pBR322载体中,分别产生pEV71(HP)和pEV71(CCA)的感染性克隆。结果表明,HP毒株的增殖速度比CCA毒株快得多。从感染性克隆中拯救出的病毒不仅在基因组序列上与其亲本毒株保持一致,而且保留了各自的生物学表型。本研究将有助于我们了解EV71的发病机制以及开发针对手足口病的新型疫苗。