A highly sensitive and specific multiplex RT-PCR to detect foot-and-mouth disease virus in tissue and food samples

Three sets of primers to detect foot-and-mouth disease virus (FMDV) using multiplex RT-PCR were designed based on several reference nucleotide sequences, and their reaction conditions were determined. By testing ten-fold serial dilutions of FMDV, the sensitivity of multiplex RT-PCR is 100 times higher than conventional RT-PCR. Meanwhile, its specificity was confirmed compared with other related vesicular disease viruses. Furthermore, 30 field samples from different animals were tested, and the results supported the method's potential applications in routine veterinary quarantine and epidemic surveillance of FMDV.