The ability of restriction enzymes to differentiate between FMD virus isolates in Egypt

Abstract

Direct detection of FMD virus from infected bovine tissues was investigated using a polymerase chain reaction (PCR) technique, two primers were constructed from consensus sequence data within the genomic region coding for the viral RNA polymerase which shows a high degree of conservation among the seven viral RNA (FMDV) serotypes.

Another two primers flanking a 372 bp in the highly variable region of the VP1 genome was constructed and used in the PCR technique for VP1 region.

The PCR products were sized by agarose gel electrophoresis, sequenced and restriction mapping were confirmed to each isolate and further examined by digestion with Asul , Fokl and Bgll confirming the predicted internal restriction enzyme site.

All isolates were amplified in a few hours and the PCR product tested positive were subjected to restriction digestion to detect the variation between different isolates , the use of PCR technique as a highly specific and rapid technique followed by restriction digestion could enabled us to differentiate between different isolates and put a key

table for further investigation to any cases appeared in the field.