There are currently five main chemistries used for the detection of PCR product during real-time PCR. These are the DNA-binding fluorophores, the 5 endonuclease, adjacent linear and hairpin oligoprobes, and the selffluorescing amplicons [156] . This approach is a highly sensitive technique enabling simultaneous amplification and quantification of specific nucleic acid sequences. In addition to enhanced sensitivity, the benefits of real-time PCR assays over conventional endpoint detection methods include their large dynamic range, a reduced risk of cross-contamination, an ability to be scaled up for high-throughput applications and the potential for accurate target quantification [158] [159] [160] . Real-time PCR assays recommended by the World Organization for animal health (OIE) for detection of FMDV incorporate universal primers and fluorescent-labeled probes that recognized conserved region within the 5 UTR [39] or conserved gene regions within the RNA-dependent RNA polymerase gene (3Dpol) [161] . The use of a specific probe facilitates an increase in specificity compared to conventional agarose-gel-based PCR assays [162, 163] .