Table 3. Rescue of multiple strains of FMDV from RNA samples isolated from bovine tongue epithelium from suspected cases of FMD in Afghanistan and Pakistan.
| Virus sample and serotype(from VP1 sequencing) | Real time RT-PCR (Ct)(5′UTR) | VirusRescued | Serotype O Ag ELISA (OD) | Serotype AAg ELISA(OD) |
| A/KUN/AFG/L629/2009 | 16.6 | - | ||
| Asia-1/BAM/AFG/L640/2009 | 16.9 | - | ||
| Asia-1/BAM/AFG/L641/2009 | 16.4 | - | ||
| Asia-1/BAM/AFG/L643/2009 | 15.4 | - | ||
| O/SIN/PAK/L693/2009 | 21.8 | - | ||
| O/PUN/PAK/L1347/2008 | 20.2 | - | ||
| A/PUN/PAK/1354/2009 | 13.8 | - | ||
| A/PUN/PAK/1355/2009 | 21.0 | - | ||
| O/PUN/PAK/L1358/2008 | 17.9 | - | ||
| A/ISL/PAK/L1411/2009 | 19.2 | - | ||
| O/ISL/PAK/L1413/2009 | 19.0 | + | 1.56 | 0.09 |
| A/BAL/AFG/L1430/2009 | 18.7 | - | ||
| A/SAR/AFG/L1435/2009 | 18.2 | - | ||
| A/HIR/AFG/L1483/2009 | 13.7 | + | 0.15 | 1.53 |
| A/HIR/AFG/L1485/2009 | 14.4 | + | 0.03 | * 0.31 |
| A/KAP/AFG/L1491/2009 | 15.1 | - | ||
| A/KUN/AFG/L1495/2009 | 16.3 | + | 0.21 | 1.75 |
The presence of FMDV RNA was determined using qRT-PCR assays, the serotype was determined from sequencing of amplicons including the VP1 coding sequence and then the RNA was introduced into BHK cells by electroporation as described in Material and Methods. When virus was successfully rescued, as determined by the generation of CPE (indicated by (+)), the harvested viruses (diluted 1∶25) were assayed for the presence of FMDV antigen using serotype specific ELISAs.
*Incomplete CPE was apparent in this sample.