Background:
Classical swine fever (CSF), caused by a Pestivirus, is one of the leading causes of economic losses in pig production. No indigenous real-time assay kit has been available so far for the detection of CSF in porcine clinical samples:
Technology Details:
The goal of this study was to verify a hydrolysis-based qPCR for detecting CSF in swine tissues. The national and international traffic of live pigs and their by-products poses a concern to CSF-free areas or countries. Many symptoms are not solely connected with the disease and may vary according to the virus strain, age, and health state of the animals; thus, laboratory identification of CSF is critical. Such identification must be quick and precise, especially in nations with established eradication initiatives, because quick diagnosis reduces the potential for transmission for uninfected herds and so avoids disease spread. The gold standard CSF diagnostic of virus isolation can be challenging, especially given the biosecurity hazards associated with handling live infectious organisms. Furthermore, serological tests used to diagnose CSF may produce contradictory findings since antibodies for other Pestivirus-caused diseases, such as bovine viral diarrhea (BVDV) and border disease, can cross-react with the CSF virus. Here, we developed a qPCR-based assay for specific detection of CSF in clinical samples. The developed assay can be able to detect the CSF viral load in clinical samples within 30 minutes (post nucleic acid extraction). Disclosed herein are the primers and probe sequences specific for the detection of CSFV in clinical samples comprising of a) forward primer (Seq 1), b) reverse primer (Seq 2), c) probe sequence (Seq 3) comprising bases labelled with XX (at 5' end) & YY (at 3' end). Further, the disclosure is a process for quantitative detection of CSFV copy numbers in clinical samples within 30 minutes using seq 1-3. The assay is specific for CSFV without any cross-reactivity with other swine viral pathogens. The detection limit of the assay was 50fg copies of standard plasmid DNA containing CSFV-specific genes.