A real-time reverse transcription-PCR (rRT-PCR) for the detection of the foot-and-mouth disease virus (FM DV) in samples of archival isolates of FMDV serotypes O, A, C, SAT1-3, and Asia 1 was described. A primer set that targets the IRES region of the FMDV genome and TaqMan probe specific for a highly-conserved region in FMDV RNA IRES region was used. The assay detected the viral RNA in all tested archival serotypes of FMDV. Swine vesicular disease virus (SVDV) and bluetongue virus (BTV) RNA as well as RNA isolated from epithelium taken from healthy uninfected calf (negative control) were undetectable in the test. The detection limit of rRT-PCR was quantified for 1 TCID₅₀. The test can provide quantitative as well as qualitative information and is more sensitive and much faster to perform than the conventional RT-PCR. It can be used in large-scale screening because of its ability to simultaneous analysis up to 96 samples per run. The applied rRT-PCR is accurate, specific, and reliable technique for the detection of FMDV in biological samples. Therefore it is seen as a valuable tool to complement the routine diagnostic procedure for FMD virus diagnosis.
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