Classical swine fever (CSF) is an economically devastating disease of pigs. Instrumental to the control of CSF is a well-characterized sensitive assay that can deliver a rapid and accurate diagnosis before the onset of clinical signs. With this objective, a real-time fluorogenic-probe hydrolysis (TaqMan) reverse transcription-polymerase chain reaction (RT-PCR) assay was developed for rapid and specific detection of classical swine fever virus (CSFV) and applied on samples derived from infected slaughtered pigs. A pair of PCR primers targeting 5’ -non-coding region (CSFL1 and CSFR1) in conjunction with a CSFV-specific fluorogenic probe (CSFP1) was designed and assessed in real-time PCR. During PCR, when the target of interest was present, the CSFV specific FAM-labeled TaqMan probe annealed to the amplicon between the forward and reverse primers and was subsequently cleaved via the 5¢-3¢ exonuclease activity of the DNA polymerase resulting in the release of the fluorescent reporter dye. This assay was found to be rapid and strain-specific for CSFV detection.
STANDARDIZATION AND VALIDATION OF QUALITATIVE REAL TIME RT-PCR FOR DETECTION OF CLASSICAL SWINE FEVER VIRUS
