Engineering infectious foot-and-mouth disease virus in vivo from a full-length genomic cDNA clone of the A/AKT/58 strain

Abstract

Two full-length genomic cDNA clones, pTA/FMDV and pCA/FMDV, were constructed that contained three point-mutants [A174G and A308G (not present in pTA/FMDV); T1029G] in the genome compared with the wild type A/AKT/58 strain of foot-and-mouth disease virus. These two viruses were rescued by co-transfection of pCA/FMDV with pCT7RNAP, which can express T7 RNA polymerase in BHK-21 cell-lines, or by transfection of the in vitro transcribed RNA. Their biological properties were analyzed for their antigenicity, virulence in suckling-mice (LD50) and growth kinetics in BHK-21 cells. The in vivo rescued viruses showed high pathogenicity for 3-day-old unweaned mice (LD50=10(-7.5)). However, the in vitro transcribed RNA derived from pTA/FMDV had lower pathogenicity for suckling-mice (LD50=10(-6)), and the in vivo transcribed RNA recovered from pCA/FMDV co-transfected with pCT7RNAP showed no significant differences from the wild type virus. These data showed that recovery of the infectious foot-and-mouth disease virus directly from the use of in vivo techniques was better than from in vitro methods. Furthermore, the reverse genetic procedure technique was simplified to a faster one-step procedure based on co-transfection with pCT7RNAP. These results suggest that in vivo RNA transcripts may be more valuable for engineering recombinant foot-and-mouth disease virus than in vitro RNA transcripts, and may contribute to further understanding of the biological properties, such as replication, maturation and quasispecies, of the foot-and-mouth disease virus.

Effects of two amino acid substitutions in the capsid proteins on the interaction of two cell-adapted PanAsia-1 strains of foot-and-mouth disease virus serotype O with heparan sulfate receptor

“…The eukaryotic expression plasmid pCT7RNAP can express T7 RNA polymerase (T7 RNAP) in BHK-21 cells [41]. Plasmid pOFS containing the full-length infectious cDNA of FMDV O/HN/CHA/93 (Cathay topotype) under the control of the bacteriophage T7 promoter was constructed by Li et al [42].…”

Development and validation of a prokaryotically expressed foot-and-mouth disease virus non-structural protein 2C'3AB-based immunochromatographic strip to differentiate between infected and vaccinated animals

“…The non-structural proteins (NSPs) are conserved amongst the seven serotypes[1,2]. FMDV serotypes O, Asia 1 and A have circulated separately in China during the last fifty years[3-6], and eliminating infected animals and vaccination are the main policies to prevent and control FMD. Antibodies to NSPs exist in infected animals, and were utilized to differentiate between infected and vaccinated animals.…”

Spatial pattern of foot-and-mouth disease in animals in China, 2010–2016

“…In China, FMD was first reported in 1958 ( Bai et al, 2011 ). In that same year, serotypes O and A isolates were collected from the Xinjiang Uygur Autonomous Region ( Bai et al, 2009 ; Li et al, 2007 ) and serotype Asia 1 isolate was collected from Yunnan province ( Du et al, 2007 ). In the following years, a series of effective measures were undertaken to prevent the spread of FMD.…”