Indian Journal of Animal Research

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Research Article

Indian Journal of Animal Research, volume 50 issue 6 (december 2016) : 979-982

Development and application of real-time TaqMan RT-PCR assay for improved detection of classical swine fever virus in slaughtered pigs 

M. Rout*, G. Saikumar1
1<p>Division of Pathology,&nbsp;Indian Veterinary Research Institute, Izatnagar - 243 122, India.</p>
Cite article:- Rout* M., Saikumar1 G. (2016). Development and application of real-time TaqMan RT-PCR assay for improved detection of classical swine fever virus in slaughtered pigs . Indian Journal of Animal Research. 50(6): 979-982. doi: 10.18805/ijar.v0iOF.6667.

ABSTRACT

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. 

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