Detection of foot-and-mouth disease virus from culture and clinical samples by reverse transcription-PCR coupled to restriction enzyme and sequence analysis.

A reverse transcription-PCR (RT-PCR) method is presented for the highly sensitive and specific detection of foot-and-mouth disease virus (FMDV). A primer pair flanking a region of the viral polymerase gene (3D) corresponding to the C-terminus of the protein was designed and a single step RT-PCR reaction was developed. The assay allowed the detection of viral RNA from a variety of animal samples and from a wide range of FMDV isolates of different origins and serotypes.

IRES-driven translation is stimulated separately by the FMDV 3'-NCR and poly(A) sequences.

The 3' end region of foot-and-mouth disease virus (FMDV) consists of two distinct elements, a 90 nt untranslated region (3'-NCR) and a poly(A) tract. Removal of either the poly(A) tract or both the 3'-NCR and the poly(A) tract abrogated infectivity in susceptible cells in the context of a full-length cDNA clone. We have addressed the question of whether the impairment of RNA infectivity is related to defects at the translation level using a double approach.